w*i
2 楼
这些声音，我们在职场中常常听到，其实有些也来自自己的内心
“凭什么他一个刚来的新员工，工资竟然比我这个入职5年的都高？”
“这个项目明明是我完成的，凭什么功劳都是他的了？”
“明明是我在天天加班加点，凭什么升职的却是她？”
醒醒吧，小伙伴，从你踏入职场的第一天起，就不该期待公平，更何况，你所谓的公平
，只是来自于你内心的看法罢了。
世界上本来就没有那么多“你应该”和“本来就”，职场上弱肉强食，优胜劣汰的故事
看的太多，踩着被人上位，一将功成万骨枯的故事天天在上演。
一个刚来一个月的新员工，说不定是你的老板高薪从猎头手里挖过来的人才；
这个项目明明是你完成的，但是人家会领导啊，会做报告啊，会做PPT啊，哄得老板心
花怒放，功劳不算人家的算谁的？
明明是你在加班加点，但人家下班的时间也没闲着，读书进修考证，英语说的比老外都
溜，以后接待大客户，难道就靠你的加班审批表吗？
从小到大，我看到最公平的事情，大概就是高考了，就连高考，也有不公平啊，为什么
北京的孩子可以轻易考上重点大学，山东的孩子600多分还上不了211？
世间的公平是相对的，不是绝对的，打铁还需自身硬，与其纠结抱怨为什么不公平，还
是先自己修炼修炼吧！
“凭什么他一个刚来的新员工，工资竟然比我这个入职5年的都高？”
“这个项目明明是我完成的，凭什么功劳都是他的了？”
“明明是我在天天加班加点，凭什么升职的却是她？”
醒醒吧，小伙伴，从你踏入职场的第一天起，就不该期待公平，更何况，你所谓的公平
，只是来自于你内心的看法罢了。
世界上本来就没有那么多“你应该”和“本来就”，职场上弱肉强食，优胜劣汰的故事
看的太多，踩着被人上位，一将功成万骨枯的故事天天在上演。
一个刚来一个月的新员工，说不定是你的老板高薪从猎头手里挖过来的人才；
这个项目明明是你完成的，但是人家会领导啊，会做报告啊，会做PPT啊，哄得老板心
花怒放，功劳不算人家的算谁的？
明明是你在加班加点，但人家下班的时间也没闲着，读书进修考证，英语说的比老外都
溜，以后接待大客户，难道就靠你的加班审批表吗？
从小到大，我看到最公平的事情，大概就是高考了，就连高考，也有不公平啊，为什么
北京的孩子可以轻易考上重点大学，山东的孩子600多分还上不了211？
世间的公平是相对的，不是绝对的，打铁还需自身硬，与其纠结抱怨为什么不公平，还
是先自己修炼修炼吧！
T*y
3 楼
I want to start a new post on this question, also as a response to Yisu's
post earlier.
I also feel that the will to understand the Truth is very important, and it
is closely related to the faith.
Well, what is the will to find the Truth? What is it exactly? Is it the
belief that there must be a reason, aka, karma? Is it the faith that there
must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
a soul, or collective unconscious, or whatever it's called) that keeps
calling on us to pursue it?
Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
want an answer not by Buddhism Sutra, but by what you feel in heart, or what
you understand as of now, in your current status.
I am asking this also because I was reading that book "Conversations with
God, book 2" over the Thanksgiving break, and I watched the DVD, which might
be based on book 1 that I haven't read yet. In the DVD, Neale had
encountered a very bad car accident, lost his job, lived in a tent in a park
as being practically homeless for a long time, before he found himself a
new job, and yet this radio station (he was a DJ there) went bankrupt
without anyone telling him anything beforehand. That's when he asked God why
why why till he found the answers in his head that he just needed to write
it down on a notepad.
The key thing I want to understand more is the concept of time, and this
book explains it as being a spindle with leaves of pages, as if each page is
a time instant, and they are always there, and we can peek into the past or
the future, or essentially there's no time, or, there is only present
moment.
Then regarding where God is, this book answers it as everywhere, as What It
Is. Since it's everywhere, it's also nowhere, as any place of itself would
be a separation from other places, but it's not, it's everywhere. Also, as
mentioned by freeman08, nowhere= Now Here.
发信人: Yisu (大头教主), 信区: Wisdom
标 题: Re: Carl Jung's Near Death Experience in 1944 (zz)
发信站: BBS 未名空间站 (Thu Nov 24 11:55:10 2011, 美东)
Sorry, I missed this post..Let me take a look..the question is why do you
think that Buddha can become Budda?
En...I think firstly it because he, like anyone else, has the potential to
become a Budda. It's like a seed planted, and with certain condition, it
surely will flourish one day. According to the Budda, everyone has the seed,
and everyone will eventually be there. But speaking of the last one, why is
him, not Y is U, or not Y is me..
I think the most imporant difference between he and us is the WILL.
And also how strong the will is to know the truth, to practice, and to get
there.
The inital will is so strong that it supports practise over the countless
lifes and the practice on the way reversely make the will stronger, making
it a healhty progressing.
Those who think he has not been improved over years should check the will
first, is that strong enough. Like the exmaple I mentioned in another post,
sth like a person's will to get above the water.
I believed he has undergone much much more practices and tests than us in
all his lifes. looks like that's the reason, but that could be a result at
the same time too..
post earlier.
I also feel that the will to understand the Truth is very important, and it
is closely related to the faith.
Well, what is the will to find the Truth? What is it exactly? Is it the
belief that there must be a reason, aka, karma? Is it the faith that there
must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
a soul, or collective unconscious, or whatever it's called) that keeps
calling on us to pursue it?
Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
want an answer not by Buddhism Sutra, but by what you feel in heart, or what
you understand as of now, in your current status.
I am asking this also because I was reading that book "Conversations with
God, book 2" over the Thanksgiving break, and I watched the DVD, which might
be based on book 1 that I haven't read yet. In the DVD, Neale had
encountered a very bad car accident, lost his job, lived in a tent in a park
as being practically homeless for a long time, before he found himself a
new job, and yet this radio station (he was a DJ there) went bankrupt
without anyone telling him anything beforehand. That's when he asked God why
why why till he found the answers in his head that he just needed to write
it down on a notepad.
The key thing I want to understand more is the concept of time, and this
book explains it as being a spindle with leaves of pages, as if each page is
a time instant, and they are always there, and we can peek into the past or
the future, or essentially there's no time, or, there is only present
moment.
Then regarding where God is, this book answers it as everywhere, as What It
Is. Since it's everywhere, it's also nowhere, as any place of itself would
be a separation from other places, but it's not, it's everywhere. Also, as
mentioned by freeman08, nowhere= Now Here.
发信人: Yisu (大头教主), 信区: Wisdom
标 题: Re: Carl Jung's Near Death Experience in 1944 (zz)
发信站: BBS 未名空间站 (Thu Nov 24 11:55:10 2011, 美东)
Sorry, I missed this post..Let me take a look..the question is why do you
think that Buddha can become Budda?
En...I think firstly it because he, like anyone else, has the potential to
become a Budda. It's like a seed planted, and with certain condition, it
surely will flourish one day. According to the Budda, everyone has the seed,
and everyone will eventually be there. But speaking of the last one, why is
him, not Y is U, or not Y is me..
I think the most imporant difference between he and us is the WILL.
And also how strong the will is to know the truth, to practice, and to get
there.
The inital will is so strong that it supports practise over the countless
lifes and the practice on the way reversely make the will stronger, making
it a healhty progressing.
Those who think he has not been improved over years should check the will
first, is that strong enough. Like the exmaple I mentioned in another post,
sth like a person's will to get above the water.
I believed he has undergone much much more practices and tests than us in
all his lifes. looks like that's the reason, but that could be a result at
the same time too..
n*1
4 楼
坏了，保修的话，我是该找buy.com 还是 beach camera 还是WD?
谢谢。。。
谢谢。。。
d*0
5 楼
卖没卖货不知道，反正是搭着顺风车出了名
p*m
6 楼
刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
（只考虑非发育生物学的应用，这个做的已经没太大意思了）
small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
orter的也不错（但是看下面）
同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
处是transient expression,这个的能量我还要继续发掘研究。以后要做genetics，也就
最多停留在这个步骤了
fish前辈们怎么看？
大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
（只考虑非发育生物学的应用，这个做的已经没太大意思了）
small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
orter的也不错（但是看下面）
同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
处是transient expression,这个的能量我还要继续发掘研究。以后要做genetics，也就
最多停留在这个步骤了
fish前辈们怎么看？
a*n
7 楼
置顶最后一篇，里面有告诉你怎么查询，然后上去后你要点那个查询签证状态那个
用的是160确认号。
你这个看不出来啥，签过不签过护照都要送回中信。
【在 C*I 的大作中提到】
: 网上发现状态变为：
: “护照已从领事馆那边收回，目前正在安排运送 ”
: 代签的说。
用的是160确认号。
你这个看不出来啥，签过不签过护照都要送回中信。
【在 C*I 的大作中提到】
: 网上发现状态变为：
: “护照已从领事馆那边收回，目前正在安排运送 ”
: 代签的说。
K*k
8 楼
呵呵，你好像问不只一个问题
will, belief, intuition,
GOD(s), truth, faith, time...
估计没有一个是有标准答案的
就个人感觉友情灌点水。
“Well, what is the will to find the Truth? What is it exactly? Is it the
belief that there must be a reason, aka, karma? Is it the faith that there
must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
a soul, or collective unconscious, or whatever it's called) that keeps
calling on us to pursue it?”
不太清楚，但是圣经里的一句话，
比较符合我的感觉
...and the truth will set you free." (John 8:32)
“Do you think that there is only ONE, or only nirvana is the ultimate ONE?
I
want an answer not by Buddhism Sutra, but by what you feel in heart, or what
you understand as of now, in your current status.”
就我个人而言，GOD就一个
那就是自然界的规律。
但反映到人的意识时
会有不同的认识。
“The key thing I want to understand more is the concept of time, and this
book explains it as being a spindle with leaves of pages, as if each page is
a time instant, and they are always there, and we can peek into the past or
the future, or essentially there's no time, or, there is only present
moment.”
我对时间认识来自钟和相对论
但是我还是不懂的时间到底是什麽。呵呵
“Then regarding where God is, this book answers it as everywhere, as What
It
Is. Since it's everywhere, it's also nowhere, as any place of itself would
be a separation from other places, but it's not, it's everywhere. Also, as
mentioned by freeman08, nowhere= Now Here.”
自然规律无处不在。
所以GOD也是无处不在。
it
of
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
will, belief, intuition,
GOD(s), truth, faith, time...
估计没有一个是有标准答案的
就个人感觉友情灌点水。
“Well, what is the will to find the Truth? What is it exactly? Is it the
belief that there must be a reason, aka, karma? Is it the faith that there
must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
a soul, or collective unconscious, or whatever it's called) that keeps
calling on us to pursue it?”
不太清楚，但是圣经里的一句话，
比较符合我的感觉
...and the truth will set you free." (John 8:32)
“Do you think that there is only ONE, or only nirvana is the ultimate ONE?
I
want an answer not by Buddhism Sutra, but by what you feel in heart, or what
you understand as of now, in your current status.”
就我个人而言，GOD就一个
那就是自然界的规律。
但反映到人的意识时
会有不同的认识。
“The key thing I want to understand more is the concept of time, and this
book explains it as being a spindle with leaves of pages, as if each page is
a time instant, and they are always there, and we can peek into the past or
the future, or essentially there's no time, or, there is only present
moment.”
我对时间认识来自钟和相对论
但是我还是不懂的时间到底是什麽。呵呵
“Then regarding where God is, this book answers it as everywhere, as What
It
Is. Since it's everywhere, it's also nowhere, as any place of itself would
be a separation from other places, but it's not, it's everywhere. Also, as
mentioned by freeman08, nowhere= Now Here.”
自然规律无处不在。
所以GOD也是无处不在。
it
of
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
a*e
9 楼
WD,一个月内退货换货找bc
bc不理你可以找找buy
【在 n******1 的大作中提到】
: 坏了，保修的话，我是该找buy.com 还是 beach camera 还是WD?
: 谢谢。。。
bc不理你可以找找buy
【在 n******1 的大作中提到】
: 坏了，保修的话，我是该找buy.com 还是 beach camera 还是WD?
: 谢谢。。。
o*4
10 楼
牛逼
C*I
11 楼
你是说这个吗：https://ceac.state.gov/CEACStatTracker/Status.aspx?eQs=
WwjqOlbeRYzCYubaSQI+RA==
查了，护照都在路上了可还是： Administrative Processing
WwjqOlbeRYzCYubaSQI+RA==
查了，护照都在路上了可还是： Administrative Processing
Y*u
12 楼
哈，为了能表达清楚一些，我还是用中文吧，
这么多问题，也不知道该从哪里开始说，就说到哪里是哪里吧。。反正都是个人看法.
佛是悟了的众生，而众生是未悟得佛。 可是佛为什么能悟呢，却也不是和众生豪无关
系，佛的力量来自众生。 为什么这么说呢？
还是举了例子吧。 一杯水，你拿起来摇一摇，有一滴水掉了出来。 我们来仔细考察一
下这一滴掉出来的水珠。 是什么让他掉了出来，而不是别的水珠？ 摇动是一个因，还
有什么呢？
是否可以说，没有别的水珠，在相同的摇动下，它也可以出来呢？ 显然，其它的水珠
的存在是这个水珠出来的另一原因(反作用力)。唯一不同的是水珠没有人一样的意识。
佛的觉悟过程也是类似的，在因缘条件满足下，肯定有一个人要成佛，但这是统一作用
的结果，即是个人的愿望和实践，又是众生的愿望和实践。
佛在忍辱仙人的时候，被人肢解，反对那个人说，以后我成佛，先来度你。 什么意思
？ 因为正是这个人帮了他大忙。 佛法在世间，不离世间觉。 世间无常是苦，觉者反
转为菩提。 佛以众生心为心，众生有多苦，佛就有多少慈悲。 所以我们问佛的力量和
智慧从哪里来，我觉得就是众生。
那么这个世界的本质是不是一个。 与神对话是一本很有意思的书。我认同其中很多道
理。比如说第一本中提到的神圣二分法。 这一点在中国传统宗教中，尤其是道教，反
映的比较多。道教是隐晦隐晦地讲，而这本书就是明明白白地说。
是，这本书的确很多地方提到了唯一的存在。我个人觉得可能是语言文字的效果， 那
可能并不是他想要表达的。 襌宗有以个公案讲的是有益个和尚问赵州：万法归一，一
归何所？赵州曰：“老僧在青州，作得一领布衫，重七斤。。。
这个公案是在打破这个和尚对一的执着。 每一个执着， 不管是对一也好，对万也好
，都是应该放下的。 一又在哪里呢？ 一不就是万法吗。。道家讲道生一，一生。。。
， 一从道来， 但是道却非一非二。 道非一二，却以一二的形式来展现。
所以我不认为有这么“一个”，因为我希望能真正见到这么“一个”。
it
of
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
这么多问题，也不知道该从哪里开始说，就说到哪里是哪里吧。。反正都是个人看法.
佛是悟了的众生，而众生是未悟得佛。 可是佛为什么能悟呢，却也不是和众生豪无关
系，佛的力量来自众生。 为什么这么说呢？
还是举了例子吧。 一杯水，你拿起来摇一摇，有一滴水掉了出来。 我们来仔细考察一
下这一滴掉出来的水珠。 是什么让他掉了出来，而不是别的水珠？ 摇动是一个因，还
有什么呢？
是否可以说，没有别的水珠，在相同的摇动下，它也可以出来呢？ 显然，其它的水珠
的存在是这个水珠出来的另一原因(反作用力)。唯一不同的是水珠没有人一样的意识。
佛的觉悟过程也是类似的，在因缘条件满足下，肯定有一个人要成佛，但这是统一作用
的结果，即是个人的愿望和实践，又是众生的愿望和实践。
佛在忍辱仙人的时候，被人肢解，反对那个人说，以后我成佛，先来度你。 什么意思
？ 因为正是这个人帮了他大忙。 佛法在世间，不离世间觉。 世间无常是苦，觉者反
转为菩提。 佛以众生心为心，众生有多苦，佛就有多少慈悲。 所以我们问佛的力量和
智慧从哪里来，我觉得就是众生。
那么这个世界的本质是不是一个。 与神对话是一本很有意思的书。我认同其中很多道
理。比如说第一本中提到的神圣二分法。 这一点在中国传统宗教中，尤其是道教，反
映的比较多。道教是隐晦隐晦地讲，而这本书就是明明白白地说。
是，这本书的确很多地方提到了唯一的存在。我个人觉得可能是语言文字的效果， 那
可能并不是他想要表达的。 襌宗有以个公案讲的是有益个和尚问赵州：万法归一，一
归何所？赵州曰：“老僧在青州，作得一领布衫，重七斤。。。
这个公案是在打破这个和尚对一的执着。 每一个执着， 不管是对一也好，对万也好
，都是应该放下的。 一又在哪里呢？ 一不就是万法吗。。道家讲道生一，一生。。。
， 一从道来， 但是道却非一非二。 道非一二，却以一二的形式来展现。
所以我不认为有这么“一个”，因为我希望能真正见到这么“一个”。
it
of
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
n*1
13 楼
过了一个月了。。我找WD吧。。谢谢～
【在 a***e 的大作中提到】
: WD,一个月内退货换货找bc
: bc不理你可以找找buy
【在 a***e 的大作中提到】
: WD,一个月内退货换货找bc
: bc不理你可以找找buy
p*p
14 楼
凳子。
a*n
15 楼
护照那个状态我没有研究过，不过签证是这个状态说明还在处理。等1-2工作日
看结果。
以前经验是issued是签过，改面签是no status
【在 C*I 的大作中提到】
: 你是说这个吗：https://ceac.state.gov/CEACStatTracker/Status.aspx?eQs=
: WwjqOlbeRYzCYubaSQI+RA==
: 查了，护照都在路上了可还是： Administrative Processing
看结果。
以前经验是issued是签过，改面签是no status
【在 C*I 的大作中提到】
: 你是说这个吗：https://ceac.state.gov/CEACStatTracker/Status.aspx?eQs=
: WwjqOlbeRYzCYubaSQI+RA==
: 查了，护照都在路上了可还是： Administrative Processing
N*n
16 楼
Interesting discussion and I want to give you my thoughts.
The "ONE" or nirvana, or whatever you want to call it, is within you. It is
your true nature. So what it our true nature? We all want to be happy, we
all love our our parents, we all want to be worry free, we all want to do
things effortlessly and fearlessly, etc., this is our true nature - the
state of happiness, love and kindness, fearless and no worry. This state is
born with us, it is our true nature. Nothing can change this state, and this
state does not depend on any outside support. This is the truth. When we
don't recognize our true nature, this is called "无明". Buddhism is a path
to discover our true nature and be with our true nature so that we don't
have suffering.
it
of
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
The "ONE" or nirvana, or whatever you want to call it, is within you. It is
your true nature. So what it our true nature? We all want to be happy, we
all love our our parents, we all want to be worry free, we all want to do
things effortlessly and fearlessly, etc., this is our true nature - the
state of happiness, love and kindness, fearless and no worry. This state is
born with us, it is our true nature. Nothing can change this state, and this
state does not depend on any outside support. This is the truth. When we
don't recognize our true nature, this is called "无明". Buddhism is a path
to discover our true nature and be with our true nature so that we don't
have suffering.
it
of
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
M*n
17 楼
俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
developmental biology is pretty much over. don't waster your time on it.
pushing/shaking flies is good for beginners to learn genetics.
my 2 cents.
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
developmental biology is pretty much over. don't waster your time on it.
pushing/shaking flies is good for beginners to learn genetics.
my 2 cents.
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
C*I
18 楼
谢谢了。good luck to my parents. 下次直接办绿卡，现在签证太麻烦了。
T*y
19 楼
hehe, Zan 友情灌水. I agree that everyone may have his/her own
interpretation on this.
【在 K*****k 的大作中提到】
: 呵呵，你好像问不只一个问题
: will, belief, intuition,
: GOD(s), truth, faith, time...
: 估计没有一个是有标准答案的
: 就个人感觉友情灌点水。
: “Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?”
interpretation on this.
【在 K*****k 的大作中提到】
: 呵呵，你好像问不只一个问题
: will, belief, intuition,
: GOD(s), truth, faith, time...
: 估计没有一个是有标准答案的
: 就个人感觉友情灌点水。
: “Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?”
p*m
20 楼
有ZFN就够了吧 为啥一定要knockout fish?
我本来就是fly person, hehe。来试一下fish水多深，不行我还弄我的fly去。。
【在 M*****n 的大作中提到】
: 俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
: 你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
: developmental biology is pretty much over. don't waster your time on it.
: pushing/shaking flies is good for beginners to learn genetics.
: my 2 cents.
:
: model
: transpare
: rep
: to
我本来就是fly person, hehe。来试一下fish水多深，不行我还弄我的fly去。。
【在 M*****n 的大作中提到】
: 俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
: 你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
: developmental biology is pretty much over. don't waster your time on it.
: pushing/shaking flies is good for beginners to learn genetics.
: my 2 cents.
:
: model
: transpare
: rep
: to
T*y
21 楼
en. It makes sense.
I tend to believe that it does not matter which way one chooses to be
connected with this ONE/Wisdom/God (whatever it's called), and religions are
especially limiting with rituals and bans. There's some discussion in this
book on how Christianity is ridiculously practiced, and it seems the same to
me in other religions. However, a good teacher is also very helpful on this
journey. Both are needed.
.佛是悟了的众生，而众生是未悟得佛。 可是佛为什么能悟呢，却也不是和众生豪无关
系，佛的力量来自众生。 为什么这么说呢？
一下这一滴掉出来的水珠。 是什么让他掉了出来，而不是别的水珠？ 摇动是一个因，
还有什么呢？是否可以说，没有别的水珠，在相同的摇动下，它也可以出来呢？ 显然
，其它的水珠的存在是这个水珠出来的另一原因(反作用力)。唯一不同的是水珠没有人
一样的意识。
【在 Y**u 的大作中提到】
: 哈，为了能表达清楚一些，我还是用中文吧，
: 这么多问题，也不知道该从哪里开始说，就说到哪里是哪里吧。。反正都是个人看法.
: 佛是悟了的众生，而众生是未悟得佛。 可是佛为什么能悟呢，却也不是和众生豪无关
: 系，佛的力量来自众生。 为什么这么说呢？
: 还是举了例子吧。 一杯水，你拿起来摇一摇，有一滴水掉了出来。 我们来仔细考察一
: 下这一滴掉出来的水珠。 是什么让他掉了出来，而不是别的水珠？ 摇动是一个因，还
: 有什么呢？
: 是否可以说，没有别的水珠，在相同的摇动下，它也可以出来呢？ 显然，其它的水珠
: 的存在是这个水珠出来的另一原因(反作用力)。唯一不同的是水珠没有人一样的意识。
: 佛的觉悟过程也是类似的，在因缘条件满足下，肯定有一个人要成佛，但这是统一作用
I tend to believe that it does not matter which way one chooses to be
connected with this ONE/Wisdom/God (whatever it's called), and religions are
especially limiting with rituals and bans. There's some discussion in this
book on how Christianity is ridiculously practiced, and it seems the same to
me in other religions. However, a good teacher is also very helpful on this
journey. Both are needed.
.佛是悟了的众生，而众生是未悟得佛。 可是佛为什么能悟呢，却也不是和众生豪无关
系，佛的力量来自众生。 为什么这么说呢？
一下这一滴掉出来的水珠。 是什么让他掉了出来，而不是别的水珠？ 摇动是一个因，
还有什么呢？是否可以说，没有别的水珠，在相同的摇动下，它也可以出来呢？ 显然
，其它的水珠的存在是这个水珠出来的另一原因(反作用力)。唯一不同的是水珠没有人
一样的意识。
【在 Y**u 的大作中提到】
: 哈，为了能表达清楚一些，我还是用中文吧，
: 这么多问题，也不知道该从哪里开始说，就说到哪里是哪里吧。。反正都是个人看法.
: 佛是悟了的众生，而众生是未悟得佛。 可是佛为什么能悟呢，却也不是和众生豪无关
: 系，佛的力量来自众生。 为什么这么说呢？
: 还是举了例子吧。 一杯水，你拿起来摇一摇，有一滴水掉了出来。 我们来仔细考察一
: 下这一滴掉出来的水珠。 是什么让他掉了出来，而不是别的水珠？ 摇动是一个因，还
: 有什么呢？
: 是否可以说，没有别的水珠，在相同的摇动下，它也可以出来呢？ 显然，其它的水珠
: 的存在是这个水珠出来的另一原因(反作用力)。唯一不同的是水珠没有人一样的意识。
: 佛的觉悟过程也是类似的，在因缘条件满足下，肯定有一个人要成佛，但这是统一作用
O*e
22 楼
这个很多人都在搞了吧，e.g.
http://www.genome.gov/25522120
【在 M*****n 的大作中提到】
: 俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
: 你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
: developmental biology is pretty much over. don't waster your time on it.
: pushing/shaking flies is good for beginners to learn genetics.
: my 2 cents.
:
: model
: transpare
: rep
: to
http://www.genome.gov/25522120
【在 M*****n 的大作中提到】
: 俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
: 你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
: developmental biology is pretty much over. don't waster your time on it.
: pushing/shaking flies is good for beginners to learn genetics.
: my 2 cents.
:
: model
: transpare
: rep
: to
T*y
23 楼
Thank you very much! I really appreciate it.
I also believe that every soul has that born and true nature, and every soul
is truly equal in this journey.
In Buddhism, there are different levels of awareness. That's something that
I am still pondering upon. How would one evaluate that? Although I hear that
measuring itself is also an obstacle to reaching either level. I want to
understand this evaluation more, as I want to know how would one know other
approaches, other than Buddhism, do not reach the same One? Could you share
more thoughts on this?
is your true nature. So what it our true nature? We all want to be happy, we
all love our our parents, we all want to be worry free, we all want to do
things effortlessly and fearlessly, etc., this is our true nature - the
state of happiness, love and kindness, fearless and no worry. This state is
born with us, it is our true nature. Nothing can change this state, and this
state does not depend on any outside support. This is the truth. When we
don't recognize our true nature, this is called "无明". Buddhism is a path
to discover our true nature and be with our true nature so that we don't
【在 N*******n 的大作中提到】
: Interesting discussion and I want to give you my thoughts.
: The "ONE" or nirvana, or whatever you want to call it, is within you. It is
: your true nature. So what it our true nature? We all want to be happy, we
: all love our our parents, we all want to be worry free, we all want to do
: things effortlessly and fearlessly, etc., this is our true nature - the
: state of happiness, love and kindness, fearless and no worry. This state is
: born with us, it is our true nature. Nothing can change this state, and this
: state does not depend on any outside support. This is the truth. When we
: don't recognize our true nature, this is called "无明". Buddhism is a path
: to discover our true nature and be with our true nature so that we don't
I also believe that every soul has that born and true nature, and every soul
is truly equal in this journey.
In Buddhism, there are different levels of awareness. That's something that
I am still pondering upon. How would one evaluate that? Although I hear that
measuring itself is also an obstacle to reaching either level. I want to
understand this evaluation more, as I want to know how would one know other
approaches, other than Buddhism, do not reach the same One? Could you share
more thoughts on this?
is your true nature. So what it our true nature? We all want to be happy, we
all love our our parents, we all want to be worry free, we all want to do
things effortlessly and fearlessly, etc., this is our true nature - the
state of happiness, love and kindness, fearless and no worry. This state is
born with us, it is our true nature. Nothing can change this state, and this
state does not depend on any outside support. This is the truth. When we
don't recognize our true nature, this is called "无明". Buddhism is a path
to discover our true nature and be with our true nature so that we don't
【在 N*******n 的大作中提到】
: Interesting discussion and I want to give you my thoughts.
: The "ONE" or nirvana, or whatever you want to call it, is within you. It is
: your true nature. So what it our true nature? We all want to be happy, we
: all love our our parents, we all want to be worry free, we all want to do
: things effortlessly and fearlessly, etc., this is our true nature - the
: state of happiness, love and kindness, fearless and no worry. This state is
: born with us, it is our true nature. Nothing can change this state, and this
: state does not depend on any outside support. This is the truth. When we
: don't recognize our true nature, this is called "无明". Buddhism is a path
: to discover our true nature and be with our true nature so that we don't
M*n
24 楼
ZFN不如homologous rec好。
有很多limitation.
你再读paper哦。
【在 p*****m 的大作中提到】
: 有ZFN就够了吧 为啥一定要knockout fish?
: 我本来就是fly person, hehe。来试一下fish水多深，不行我还弄我的fly去。。
有很多limitation.
你再读paper哦。
【在 p*****m 的大作中提到】
: 有ZFN就够了吧 为啥一定要knockout fish?
: 我本来就是fly person, hehe。来试一下fish水多深，不行我还弄我的fly去。。
N*n
25 楼
You are welcome. Good discussion is nice to have.
I am not sure what you mean by "levels of awareness". Can you explain?
If we give awareness a definition, I would say awareness is the ability to
see the movement in the mind. Or one can say awareness is the clearity of
the mind. It's the same meaning. The awareness is born with us and it can
cut though anything appears in the mind. As a begginer meditator, the
awareness is hidden behind the thoughts and emotions and can not be seen. As
one progresses in medition, the mind become calmer and the awareness become
clearer. As the meditator continues to progress, the awareness become so
clear, that it is realizes as the natural born wisdom. The awareness is the
same awareness, but the experience changes as the meditation progress.
As for your question regarding how could one know other approaches, other
than Buddhism, do not reach the same One, I can not say other approaches can
not reach the same level as Buddha. But I can say for sure that no other
approaches can surpass the level of Buddha. The awareness, this natural born
wisdom, is so sharp and it can cut through anything and any suffering, what
else can surpass it?
soul
that
that
other
share
we
【在 T*******y 的大作中提到】
: Thank you very much! I really appreciate it.
: I also believe that every soul has that born and true nature, and every soul
: is truly equal in this journey.
: In Buddhism, there are different levels of awareness. That's something that
: I am still pondering upon. How would one evaluate that? Although I hear that
: measuring itself is also an obstacle to reaching either level. I want to
: understand this evaluation more, as I want to know how would one know other
: approaches, other than Buddhism, do not reach the same One? Could you share
: more thoughts on this?
:
I am not sure what you mean by "levels of awareness". Can you explain?
If we give awareness a definition, I would say awareness is the ability to
see the movement in the mind. Or one can say awareness is the clearity of
the mind. It's the same meaning. The awareness is born with us and it can
cut though anything appears in the mind. As a begginer meditator, the
awareness is hidden behind the thoughts and emotions and can not be seen. As
one progresses in medition, the mind become calmer and the awareness become
clearer. As the meditator continues to progress, the awareness become so
clear, that it is realizes as the natural born wisdom. The awareness is the
same awareness, but the experience changes as the meditation progress.
As for your question regarding how could one know other approaches, other
than Buddhism, do not reach the same One, I can not say other approaches can
not reach the same level as Buddha. But I can say for sure that no other
approaches can surpass the level of Buddha. The awareness, this natural born
wisdom, is so sharp and it can cut through anything and any suffering, what
else can surpass it?
soul
that
that
other
share
we
【在 T*******y 的大作中提到】
: Thank you very much! I really appreciate it.
: I also believe that every soul has that born and true nature, and every soul
: is truly equal in this journey.
: In Buddhism, there are different levels of awareness. That's something that
: I am still pondering upon. How would one evaluate that? Although I hear that
: measuring itself is also an obstacle to reaching either level. I want to
: understand this evaluation more, as I want to know how would one know other
: approaches, other than Buddhism, do not reach the same One? Could you share
: more thoughts on this?
:
p*m
26 楼
limitation是有，但是也可以改进吧 我最近正在谈的一个合作就是弄个更好用的ZFN
analog,你觉得一定要HM么？
【在 M*****n 的大作中提到】
: ZFN不如homologous rec好。
: 有很多limitation.
: 你再读paper哦。
analog,你觉得一定要HM么？
【在 M*****n 的大作中提到】
: ZFN不如homologous rec好。
: 有很多limitation.
: 你再读paper哦。
T*y
27 楼
Thank you so much!
By different levels of awareness, I meant different levels or kinds of 解脱,
and the different levels of souls such as 佛、菩薩、天、人、阿修羅, etc. It
somehow seems to me that it's either a yes to be in that awareness or a no,
at each instant, but not something like 50% there, 90% there, etc. However,
I do have a level in mind, as how evolved a soul can be, in the sense how
often the soul can maintain that awareness. Once a soul reaches some
awareness, the soul may not stay in that awareness though. May I know what
you want to say more about this?
I liked your explanation on the awareness. I don't think that there's any
way that surpasses another, but rather all lead to the same One (or
awareness, or wisdom, etc). As I mentioned on this board earlier, Eckhart
Tolle, or Byron Katie, or many others had some profound spiritual
experiences, how do you evaluate their experiences by Buddhism standard, and
how confident are you in such evaluation?
see the movement in the mind. Or one can say awareness is the clearity of
the mind. It's the same meaning. The awareness is born with us and it can
cut though anything appears in the mind. As a begginer meditator, the
awareness is hidden behind the thoughts and emotions and can not be seen. As
one progresses in medition, the mind become calmer and the awareness become
clearer. As the meditator continues to progress, the awareness become so
clear, that it is realizes as the natural born wisdom. The awareness is the
【在 N*******n 的大作中提到】
: You are welcome. Good discussion is nice to have.
: I am not sure what you mean by "levels of awareness". Can you explain?
: If we give awareness a definition, I would say awareness is the ability to
: see the movement in the mind. Or one can say awareness is the clearity of
: the mind. It's the same meaning. The awareness is born with us and it can
: cut though anything appears in the mind. As a begginer meditator, the
: awareness is hidden behind the thoughts and emotions and can not be seen. As
: one progresses in medition, the mind become calmer and the awareness become
: clearer. As the meditator continues to progress, the awareness become so
: clear, that it is realizes as the natural born wisdom. The awareness is the
By different levels of awareness, I meant different levels or kinds of 解脱,
and the different levels of souls such as 佛、菩薩、天、人、阿修羅, etc. It
somehow seems to me that it's either a yes to be in that awareness or a no,
at each instant, but not something like 50% there, 90% there, etc. However,
I do have a level in mind, as how evolved a soul can be, in the sense how
often the soul can maintain that awareness. Once a soul reaches some
awareness, the soul may not stay in that awareness though. May I know what
you want to say more about this?
I liked your explanation on the awareness. I don't think that there's any
way that surpasses another, but rather all lead to the same One (or
awareness, or wisdom, etc). As I mentioned on this board earlier, Eckhart
Tolle, or Byron Katie, or many others had some profound spiritual
experiences, how do you evaluate their experiences by Buddhism standard, and
how confident are you in such evaluation?
see the movement in the mind. Or one can say awareness is the clearity of
the mind. It's the same meaning. The awareness is born with us and it can
cut though anything appears in the mind. As a begginer meditator, the
awareness is hidden behind the thoughts and emotions and can not be seen. As
one progresses in medition, the mind become calmer and the awareness become
clearer. As the meditator continues to progress, the awareness become so
clear, that it is realizes as the natural born wisdom. The awareness is the
【在 N*******n 的大作中提到】
: You are welcome. Good discussion is nice to have.
: I am not sure what you mean by "levels of awareness". Can you explain?
: If we give awareness a definition, I would say awareness is the ability to
: see the movement in the mind. Or one can say awareness is the clearity of
: the mind. It's the same meaning. The awareness is born with us and it can
: cut though anything appears in the mind. As a begginer meditator, the
: awareness is hidden behind the thoughts and emotions and can not be seen. As
: one progresses in medition, the mind become calmer and the awareness become
: clearer. As the meditator continues to progress, the awareness become so
: clear, that it is realizes as the natural born wisdom. The awareness is the
M*n
28 楼
retroviral insertion是老路子，但是也有limitation.
其实这就是NIH到中国去骗钱。
【在 O******e 的大作中提到】
: 这个很多人都在搞了吧，e.g.
: http://www.genome.gov/25522120
其实这就是NIH到中国去骗钱。
【在 O******e 的大作中提到】
: 这个很多人都在搞了吧，e.g.
: http://www.genome.gov/25522120
N*n
29 楼
佛、菩薩、天、人、等不同是在执着的程度上不同,awareness 上的区别是有没有确定
的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认知
的是mind, not true awareness.
我对Eckhart Tolle, Byron Katie都不了解, 没法作评价. 通常来说, 佛的境界,如果
没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去除
执着的漫漫长路上的一个起点.
脱,
It
no,
However,
【在 T*******y 的大作中提到】
: Thank you so much!
: By different levels of awareness, I meant different levels or kinds of 解脱,
: and the different levels of souls such as 佛、菩薩、天、人、阿修羅, etc. It
: somehow seems to me that it's either a yes to be in that awareness or a no,
: at each instant, but not something like 50% there, 90% there, etc. However,
: I do have a level in mind, as how evolved a soul can be, in the sense how
: often the soul can maintain that awareness. Once a soul reaches some
: awareness, the soul may not stay in that awareness though. May I know what
: you want to say more about this?
: I liked your explanation on the awareness. I don't think that there's any
的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认知
的是mind, not true awareness.
我对Eckhart Tolle, Byron Katie都不了解, 没法作评价. 通常来说, 佛的境界,如果
没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去除
执着的漫漫长路上的一个起点.
脱,
It
no,
However,
【在 T*******y 的大作中提到】
: Thank you so much!
: By different levels of awareness, I meant different levels or kinds of 解脱,
: and the different levels of souls such as 佛、菩薩、天、人、阿修羅, etc. It
: somehow seems to me that it's either a yes to be in that awareness or a no,
: at each instant, but not something like 50% there, 90% there, etc. However,
: I do have a level in mind, as how evolved a soul can be, in the sense how
: often the soul can maintain that awareness. Once a soul reaches some
: awareness, the soul may not stay in that awareness though. May I know what
: you want to say more about this?
: I liked your explanation on the awareness. I don't think that there's any
M*n
30 楼
knock-in, can you do it with ZFN?
【在 p*****m 的大作中提到】
: limitation是有，但是也可以改进吧 我最近正在谈的一个合作就是弄个更好用的ZFN
: analog,你觉得一定要HM么？
【在 p*****m 的大作中提到】
: limitation是有，但是也可以改进吧 我最近正在谈的一个合作就是弄个更好用的ZFN
: analog,你觉得一定要HM么？
T*y
31 楼
en. I see. Thank you! I think that I need to think more, and/or experience
more, to really understand this; also, not just mentally understanding it,
but knowing it.
定的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认
知的是mind, not true awareness.
果没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去
除执着的漫漫长路上的一个起点.
【在 N*******n 的大作中提到】
: 佛、菩薩、天、人、等不同是在执着的程度上不同,awareness 上的区别是有没有确定
: 的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认知
: 的是mind, not true awareness.
: 我对Eckhart Tolle, Byron Katie都不了解, 没法作评价. 通常来说, 佛的境界,如果
: 没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去除
: 执着的漫漫长路上的一个起点.
:
: 脱,
: It
: no,
more, to really understand this; also, not just mentally understanding it,
but knowing it.
定的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认
知的是mind, not true awareness.
果没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去
除执着的漫漫长路上的一个起点.
【在 N*******n 的大作中提到】
: 佛、菩薩、天、人、等不同是在执着的程度上不同,awareness 上的区别是有没有确定
: 的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认知
: 的是mind, not true awareness.
: 我对Eckhart Tolle, Byron Katie都不了解, 没法作评价. 通常来说, 佛的境界,如果
: 没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去除
: 执着的漫漫长路上的一个起点.
:
: 脱,
: It
: no,
p*m
32 楼
对的 这个就是我正在谈的改进版。。哈哈 看来你们fish people觉得这个会是个重要的
东东咯？
【在 M*****n 的大作中提到】
: knock-in, can you do it with ZFN?
东东咯？
【在 M*****n 的大作中提到】
: knock-in, can you do it with ZFN?
s*l
33 楼
其大无外 其小无内，若有所相，皆属虚妄。
CWG 我想主要是写给现在西方人看的。
it : is closely related to the faith.
belief that there must be a reason, aka, karma? Is it the faith that there :
must be a way out, aka, nirvana? Or is it the intuition (built-in essence
of: a soul, or collective unconscious, or whatever it's called) that keeps
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
CWG 我想主要是写给现在西方人看的。
it : is closely related to the faith.
belief that there must be a reason, aka, karma? Is it the faith that there :
must be a way out, aka, nirvana? Or is it the intuition (built-in essence
of: a soul, or collective unconscious, or whatever it's called) that keeps
I
【在 T*******y 的大作中提到】
: I want to start a new post on this question, also as a response to Yisu's
: post earlier.
: I also feel that the will to understand the Truth is very important, and it
: is closely related to the faith.
: Well, what is the will to find the Truth? What is it exactly? Is it the
: belief that there must be a reason, aka, karma? Is it the faith that there
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence of
: a soul, or collective unconscious, or whatever it's called) that keeps
: calling on us to pursue it?
: Do you think that there is only ONE, or only nirvana is the ultimate ONE? I
M*n
34 楼
knock-in is a plus, 但是knock-out 最重要。
现在ZFN的效率太低了， 最多20%。你如果能弄一个比较efficiet的ZFN,比如80%, 那么
knock-in有没有也无所谓。
要的
【在 p*****m 的大作中提到】
: 对的 这个就是我正在谈的改进版。。哈哈 看来你们fish people觉得这个会是个重要的
: 东东咯？
现在ZFN的效率太低了， 最多20%。你如果能弄一个比较efficiet的ZFN,比如80%, 那么
knock-in有没有也无所谓。
要的
【在 p*****m 的大作中提到】
: 对的 这个就是我正在谈的改进版。。哈哈 看来你们fish people觉得这个会是个重要的
: 东东咯？
b*d
35 楼
俺觉得不是的。俺觉得true nature 是没有高兴或者不高兴。 happiness is a fruit
但是不是truth. Truth is unconditional world.
However, 俺喜欢你这个attitude. 我希望你讲的是真的，但是感觉可能不是的。
is
is
this
【在 N*******n 的大作中提到】
: Interesting discussion and I want to give you my thoughts.
: The "ONE" or nirvana, or whatever you want to call it, is within you. It is
: your true nature. So what it our true nature? We all want to be happy, we
: all love our our parents, we all want to be worry free, we all want to do
: things effortlessly and fearlessly, etc., this is our true nature - the
: state of happiness, love and kindness, fearless and no worry. This state is
: born with us, it is our true nature. Nothing can change this state, and this
: state does not depend on any outside support. This is the truth. When we
: don't recognize our true nature, this is called "无明". Buddhism is a path
: to discover our true nature and be with our true nature so that we don't
但是不是truth. Truth is unconditional world.
However, 俺喜欢你这个attitude. 我希望你讲的是真的，但是感觉可能不是的。
is
is
this
【在 N*******n 的大作中提到】
: Interesting discussion and I want to give you my thoughts.
: The "ONE" or nirvana, or whatever you want to call it, is within you. It is
: your true nature. So what it our true nature? We all want to be happy, we
: all love our our parents, we all want to be worry free, we all want to do
: things effortlessly and fearlessly, etc., this is our true nature - the
: state of happiness, love and kindness, fearless and no worry. This state is
: born with us, it is our true nature. Nothing can change this state, and this
: state does not depend on any outside support. This is the truth. When we
: don't recognize our true nature, this is called "无明". Buddhism is a path
: to discover our true nature and be with our true nature so that we don't
p*m
36 楼
我没明白，20%已经比老鼠HM高得多得多了啊 你是想在同代用ZFN做cKO?这样才需要考
虑不加筛选的效率问题吧
【在 M*****n 的大作中提到】
: knock-in is a plus, 但是knock-out 最重要。
: 现在ZFN的效率太低了， 最多20%。你如果能弄一个比较efficiet的ZFN,比如80%, 那么
: knock-in有没有也无所谓。
:
: 要的
虑不加筛选的效率问题吧
【在 M*****n 的大作中提到】
: knock-in is a plus, 但是knock-out 最重要。
: 现在ZFN的效率太低了， 最多20%。你如果能弄一个比较efficiet的ZFN,比如80%, 那么
: knock-in有没有也无所谓。
:
: 要的
N*n
37 楼
True Nature 就是心的本性。心性广大如虚空，但心性不仅是如虚空，不是绝对的空
无所有。超越一切相对的明空与慈悲喜舍其实是不可分割的。我们凡夫的乐往往是有得
失，有对境的乐。而我们与生俱来的心性中的乐与慈是无得失，无对境的广大的乐和慈
。It is unconditional。
当然你说得也没错，你也可以说"happiness is the fruit,但是不是truth." 只是我们
看的角度不同。
fruit
【在 b**d 的大作中提到】
: 俺觉得不是的。俺觉得true nature 是没有高兴或者不高兴。 happiness is a fruit
: 但是不是truth. Truth is unconditional world.
: However, 俺喜欢你这个attitude. 我希望你讲的是真的，但是感觉可能不是的。
:
: is
: is
: this
无所有。超越一切相对的明空与慈悲喜舍其实是不可分割的。我们凡夫的乐往往是有得
失，有对境的乐。而我们与生俱来的心性中的乐与慈是无得失，无对境的广大的乐和慈
。It is unconditional。
当然你说得也没错，你也可以说"happiness is the fruit,但是不是truth." 只是我们
看的角度不同。
fruit
【在 b**d 的大作中提到】
: 俺觉得不是的。俺觉得true nature 是没有高兴或者不高兴。 happiness is a fruit
: 但是不是truth. Truth is unconditional world.
: However, 俺喜欢你这个attitude. 我希望你讲的是真的，但是感觉可能不是的。
:
: is
: is
: this
M*n
38 楼
俺对ZFN这个field知道不多,
好像作knock-in，还是要靠hom rec的， 虽然ZFN可以有所帮助。
要的
【在 p*****m 的大作中提到】
: 对的 这个就是我正在谈的改进版。。哈哈 看来你们fish people觉得这个会是个重要的
: 东东咯？
好像作knock-in，还是要靠hom rec的， 虽然ZFN可以有所帮助。
要的
【在 p*****m 的大作中提到】
: 对的 这个就是我正在谈的改进版。。哈哈 看来你们fish people觉得这个会是个重要的
: 东东咯？
J*s
39 楼
What is the 'true awareness'?
【在 N*******n 的大作中提到】
: 佛、菩薩、天、人、等不同是在执着的程度上不同,awareness 上的区别是有没有确定
: 的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认知
: 的是mind, not true awareness.
: 我对Eckhart Tolle, Byron Katie都不了解, 没法作评价. 通常来说, 佛的境界,如果
: 没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去除
: 执着的漫漫长路上的一个起点.
:
: 脱,
: It
: no,
【在 N*******n 的大作中提到】
: 佛、菩薩、天、人、等不同是在执着的程度上不同,awareness 上的区别是有没有确定
: 的认知.任何人都可以说"我有awareness","我认知了awareness",但是通常他们所认知
: 的是mind, not true awareness.
: 我对Eckhart Tolle, Byron Katie都不了解, 没法作评价. 通常来说, 佛的境界,如果
: 没有传承,没有系统学习的是很难达到的. 就算某些人真的认知了觉性,那也只是在去除
: 执着的漫漫长路上的一个起点.
:
: 脱,
: It
: no,
p*m
40 楼
是的
【在 M*****n 的大作中提到】
: 俺对ZFN这个field知道不多,
: 好像作knock-in，还是要靠hom rec的， 虽然ZFN可以有所帮助。
:
: 要的
【在 M*****n 的大作中提到】
: 俺对ZFN这个field知道不多,
: 好像作knock-in，还是要靠hom rec的， 虽然ZFN可以有所帮助。
:
: 要的
T*y
41 楼
That book is a conversation between Neale and God, and there're many
references to Christianity, indeed, but it mentions Buddhism as well. I
haven't finished reading it, so I can't say too much now.
【在 s*****l 的大作中提到】
: 其大无外 其小无内，若有所相，皆属虚妄。
: CWG 我想主要是写给现在西方人看的。
:
: it : is closely related to the faith.
: belief that there must be a reason, aka, karma? Is it the faith that there :
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence
: of: a soul, or collective unconscious, or whatever it's called) that keeps
: I
references to Christianity, indeed, but it mentions Buddhism as well. I
haven't finished reading it, so I can't say too much now.
【在 s*****l 的大作中提到】
: 其大无外 其小无内，若有所相，皆属虚妄。
: CWG 我想主要是写给现在西方人看的。
:
: it : is closely related to the faith.
: belief that there must be a reason, aka, karma? Is it the faith that there :
: must be a way out, aka, nirvana? Or is it the intuition (built-in essence
: of: a soul, or collective unconscious, or whatever it's called) that keeps
: I
M*n
42 楼
you are smart.
其实对于fish来说，有无knock-in无所谓，可以有别的途径，但是要做cKO, 就麻烦了。
你要不想想这个怎么弄吧。
【在 p*****m 的大作中提到】
: 我没明白，20%已经比老鼠HM高得多得多了啊 你是想在同代用ZFN做cKO?这样才需要考
: 虑不加筛选的效率问题吧
其实对于fish来说，有无knock-in无所谓，可以有别的途径，但是要做cKO, 就麻烦了。
你要不想想这个怎么弄吧。
【在 p*****m 的大作中提到】
: 我没明白，20%已经比老鼠HM高得多得多了啊 你是想在同代用ZFN做cKO?这样才需要考
: 虑不加筛选的效率问题吧
N*n
43 楼
True awareness happens when the awareness sees itself.
【在 J******s 的大作中提到】
: What is the 'true awareness'?
【在 J******s 的大作中提到】
: What is the 'true awareness'?
p*m
44 楼
嗯 我今天才看到个paper可以用在这个上面 哈 你觉得做出来这个会有多大用呢？对
fish people来说，我一个外人没感觉。。
了。
【在 M*****n 的大作中提到】
: you are smart.
: 其实对于fish来说，有无knock-in无所谓，可以有别的途径，但是要做cKO, 就麻烦了。
: 你要不想想这个怎么弄吧。
fish people来说，我一个外人没感觉。。
了。
【在 M*****n 的大作中提到】
: you are smart.
: 其实对于fish来说，有无knock-in无所谓，可以有别的途径，但是要做cKO, 就麻烦了。
: 你要不想想这个怎么弄吧。
J*s
45 楼
So how could awareness see itself when the subject and object are same?
【在 N*******n 的大作中提到】
: True awareness happens when the awareness sees itself.
【在 N*******n 的大作中提到】
: True awareness happens when the awareness sees itself.
M*n
46 楼
really? which paper?
I am planning to hire a postdoc to work on that. maybe it is too late.
【在 p*****m 的大作中提到】
: 嗯 我今天才看到个paper可以用在这个上面 哈 你觉得做出来这个会有多大用呢？对
: fish people来说，我一个外人没感觉。。
:
: 了。
I am planning to hire a postdoc to work on that. maybe it is too late.
【在 p*****m 的大作中提到】
: 嗯 我今天才看到个paper可以用在这个上面 哈 你觉得做出来这个会有多大用呢？对
: fish people来说，我一个外人没感觉。。
:
: 了。
N*n
47 楼
The awareness can see itself because there is no subject and no object to
block it.
【在 J******s 的大作中提到】
: So how could awareness see itself when the subject and object are same?
block it.
【在 J******s 的大作中提到】
: So how could awareness see itself when the subject and object are same?
s*y
48 楼
鱼的好处就是身体透明，而且发育过程都在体外吧
要是什么时候做老鼠的搞出了人造老鼠子宫，你们做鱼的就可以去摆摊卖鱼了。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
要是什么时候做老鼠的搞出了人造老鼠子宫，你们做鱼的就可以去摆摊卖鱼了。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
J*s
49 楼
I mean in your sentence 'the awareness can see itself' here subject is
awareness, and object is itself, how could the same subject see the same
object?
I am not asking the reason 'there is no subject and no object', even this is
the reason, I can't see the logical correctness of the causality.
Or can you give a clearer answer? :-)
【在 N*******n 的大作中提到】
: The awareness can see itself because there is no subject and no object to
: block it.
awareness, and object is itself, how could the same subject see the same
object?
I am not asking the reason 'there is no subject and no object', even this is
the reason, I can't see the logical correctness of the causality.
Or can you give a clearer answer? :-)
【在 N*******n 的大作中提到】
: The awareness can see itself because there is no subject and no object to
: block it.
M*n
50 楼
p同学是push fly的。
【在 s******y 的大作中提到】
: 鱼的好处就是身体透明，而且发育过程都在体外吧
: 要是什么时候做老鼠的搞出了人造老鼠子宫，你们做鱼的就可以去摆摊卖鱼了。
:
: model
: transpare
: rep
: to
: 很多
: serious的
: 大好
【在 s******y 的大作中提到】
: 鱼的好处就是身体透明，而且发育过程都在体外吧
: 要是什么时候做老鼠的搞出了人造老鼠子宫，你们做鱼的就可以去摆摊卖鱼了。
:
: model
: transpare
: rep
: to
: 很多
: serious的
: 大好
N*n
51 楼
Awareness has three special properties: emptiness, clearity and ability to
know. Ability to know I mean it can know what your are thinking or what you
are doing. We are living beings because we all have this natural ability to
know. Empty means it has no shape or color or meaning, and it is not a
subject or object. You can't even describe it. The moment you describe it,
it becomes an object and it not true anymore. Normally because of strong
grasping by the mind, our awareness is covered by the thoughts and emotions.
But awareness is always there.
Words are pretty much reach its limit here. 你可以去看看禅宗祖师的开示.
is
【在 J******s 的大作中提到】
: I mean in your sentence 'the awareness can see itself' here subject is
: awareness, and object is itself, how could the same subject see the same
: object?
: I am not asking the reason 'there is no subject and no object', even this is
: the reason, I can't see the logical correctness of the causality.
: Or can you give a clearer answer? :-)
know. Ability to know I mean it can know what your are thinking or what you
are doing. We are living beings because we all have this natural ability to
know. Empty means it has no shape or color or meaning, and it is not a
subject or object. You can't even describe it. The moment you describe it,
it becomes an object and it not true anymore. Normally because of strong
grasping by the mind, our awareness is covered by the thoughts and emotions.
But awareness is always there.
Words are pretty much reach its limit here. 你可以去看看禅宗祖师的开示.
is
【在 J******s 的大作中提到】
: I mean in your sentence 'the awareness can see itself' here subject is
: awareness, and object is itself, how could the same subject see the same
: object?
: I am not asking the reason 'there is no subject and no object', even this is
: the reason, I can't see the logical correctness of the causality.
: Or can you give a clearer answer? :-)
s*y
52 楼
偶知道，他现在不是想去做鱼么，所以我要给他浇点冷水。
鱼还有一个坏事，就是很多抗体都没有卖的。很多实验室都是索性自己做一个。
幸好据说互相之间不是很难借到。但是万一碰到不耐斯的人，就麻烦了。
【在 M*****n 的大作中提到】
: p同学是push fly的。
鱼还有一个坏事，就是很多抗体都没有卖的。很多实验室都是索性自己做一个。
幸好据说互相之间不是很难借到。但是万一碰到不耐斯的人，就麻烦了。
【在 M*****n 的大作中提到】
: p同学是push fly的。
N*n
53 楼
I was busy at work this morning and did not have time to type an example.
Now I have a little bit of time and I will try to explain to you with an
example. Suppose you are in a room. When you do not see anybody else in the
room, you know you are the only one in the room. This same situation can be
applied to awareness. Suppose this room has light - awareness is like the
light. Let's say there is nothing in the room except light. The room is
totally empty. Remember the awareness has the ability to know. Normally the
awareness is always busy knowing all the things happening in the room. Now
the room is totally empty. The awareness still try to know what's happening
in the room - try to know. What awareness sees? Only emptiness and clarity (
light), right? What is emptiness and light (clarity)? - Awareness. The
awareness is trying to know what's going on the room, instead it only see
the emptiness and clarity - which is itself. Now let's replace the room with
mind. Mind is just like a busy room, normally have lots of stuff going. But
when the mind is empty, what awareness sees? - Only emptiness and clarity.
This is one way awareness can see itself. That's why 早期襌宗提唱“无念为宗
”。
Without knowing your background on the path of Buddhism, I hope this is not
too strange or too difficult to you. But this is in deed difficult to
understand and also difficult to explain. The awareness has the ability to
know itself. This one of properties of awareness. This is the key point.
is
【在 J******s 的大作中提到】
: I mean in your sentence 'the awareness can see itself' here subject is
: awareness, and object is itself, how could the same subject see the same
: object?
: I am not asking the reason 'there is no subject and no object', even this is
: the reason, I can't see the logical correctness of the causality.
: Or can you give a clearer answer? :-)
Now I have a little bit of time and I will try to explain to you with an
example. Suppose you are in a room. When you do not see anybody else in the
room, you know you are the only one in the room. This same situation can be
applied to awareness. Suppose this room has light - awareness is like the
light. Let's say there is nothing in the room except light. The room is
totally empty. Remember the awareness has the ability to know. Normally the
awareness is always busy knowing all the things happening in the room. Now
the room is totally empty. The awareness still try to know what's happening
in the room - try to know. What awareness sees? Only emptiness and clarity (
light), right? What is emptiness and light (clarity)? - Awareness. The
awareness is trying to know what's going on the room, instead it only see
the emptiness and clarity - which is itself. Now let's replace the room with
mind. Mind is just like a busy room, normally have lots of stuff going. But
when the mind is empty, what awareness sees? - Only emptiness and clarity.
This is one way awareness can see itself. That's why 早期襌宗提唱“无念为宗
”。
Without knowing your background on the path of Buddhism, I hope this is not
too strange or too difficult to you. But this is in deed difficult to
understand and also difficult to explain. The awareness has the ability to
know itself. This one of properties of awareness. This is the key point.
is
【在 J******s 的大作中提到】
: I mean in your sentence 'the awareness can see itself' here subject is
: awareness, and object is itself, how could the same subject see the same
: object?
: I am not asking the reason 'there is no subject and no object', even this is
: the reason, I can't see the logical correctness of the causality.
: Or can you give a clearer answer? :-)
M*n
54 楼
这个市场太小。
以前有个公司打算专门作fish 的抗体，不过不知道现在怎么样了。
【在 s******y 的大作中提到】
: 偶知道，他现在不是想去做鱼么，所以我要给他浇点冷水。
: 鱼还有一个坏事，就是很多抗体都没有卖的。很多实验室都是索性自己做一个。
: 幸好据说互相之间不是很难借到。但是万一碰到不耐斯的人，就麻烦了。
以前有个公司打算专门作fish 的抗体，不过不知道现在怎么样了。
【在 s******y 的大作中提到】
: 偶知道，他现在不是想去做鱼么，所以我要给他浇点冷水。
: 鱼还有一个坏事，就是很多抗体都没有卖的。很多实验室都是索性自己做一个。
: 幸好据说互相之间不是很难借到。但是万一碰到不耐斯的人，就麻烦了。
J*s
55 楼
I was busy at work this morning and did not have time to type an example.
Now I have a little bit of time and I will try to explain to you with an
example. Suppose you are in a room. When you do not see anybody else in the
room, you know you are the only one in the room. This same situation can be
applied to awareness. Suppose this room has light - awareness is like the
light. Let's say there is nothing in the room except light. The room is
totally empty. Remember the awareness has the ability to know. Normally the
awareness is always busy knowing all the things happening in the room. Now
the room is totally empty. The awareness still try to know what's happening
in the room - try to know. What awareness sees? Only emptiness and clarity (
light), right? What is emptiness and light (clarity)? - Awareness. The
awareness is trying to know what's going on the room, instead it only see
the emptiness and clarity - which is itself. Now let's replace the room with
mind. Mind is just like a busy room, normally have lots of stuff going. But
when the mind is empty, what awareness sees? - Only emptiness and clarity.
This is one way awareness can see itself. That's why 早期襌宗提唱“无念为宗
”。
-----------
First I should say thanks a lot for your detailed reply. :-)
Why does the awareness have the ability to know the room of mind is empty?
How to let the room of mind be empty?
How could the mind be empty?
When the mind is empty, could the awareness still work?
What is the physiological basis of our busy mind and the awareness?
Do they need similar mechanisms to run?
“无念”?
How to perform “无念”?
How to judge the awareness is the true awareness?
Could you give a scientific definition or description of the awareness?
Waiting for more input from you. Thanks!
Now I have a little bit of time and I will try to explain to you with an
example. Suppose you are in a room. When you do not see anybody else in the
room, you know you are the only one in the room. This same situation can be
applied to awareness. Suppose this room has light - awareness is like the
light. Let's say there is nothing in the room except light. The room is
totally empty. Remember the awareness has the ability to know. Normally the
awareness is always busy knowing all the things happening in the room. Now
the room is totally empty. The awareness still try to know what's happening
in the room - try to know. What awareness sees? Only emptiness and clarity (
light), right? What is emptiness and light (clarity)? - Awareness. The
awareness is trying to know what's going on the room, instead it only see
the emptiness and clarity - which is itself. Now let's replace the room with
mind. Mind is just like a busy room, normally have lots of stuff going. But
when the mind is empty, what awareness sees? - Only emptiness and clarity.
This is one way awareness can see itself. That's why 早期襌宗提唱“无念为宗
”。
-----------
First I should say thanks a lot for your detailed reply. :-)
Why does the awareness have the ability to know the room of mind is empty?
How to let the room of mind be empty?
How could the mind be empty?
When the mind is empty, could the awareness still work?
What is the physiological basis of our busy mind and the awareness?
Do they need similar mechanisms to run?
“无念”?
How to perform “无念”?
How to judge the awareness is the true awareness?
Could you give a scientific definition or description of the awareness?
Waiting for more input from you. Thanks!
a*k
56 楼
anaspec and GeneTex got hundreds antibodies for fish. Better than nothing.
【在 M*****n 的大作中提到】
: 这个市场太小。
: 以前有个公司打算专门作fish 的抗体，不过不知道现在怎么样了。
【在 M*****n 的大作中提到】
: 这个市场太小。
: 以前有个公司打算专门作fish 的抗体，不过不知道现在怎么样了。
b*d
57 楼
虽然看不懂，但是俺觉得蛮有道理的。 俺的脑海里一直有这么一副画面，就是一具肉
体一动不动，然后灵魂（awareness 再加点其他东东）一附上去，他就活啦。
Our physical body is not really ours, 实在只是一个physical being. awareness
是我们吗？我也不知道。 但是感觉这个话题很有意思。 纯属闲聊了。
【在 N*******n 的大作中提到】
: True awareness happens when the awareness sees itself.
体一动不动，然后灵魂（awareness 再加点其他东东）一附上去，他就活啦。
Our physical body is not really ours, 实在只是一个physical being. awareness
是我们吗？我也不知道。 但是感觉这个话题很有意思。 纯属闲聊了。
【在 N*******n 的大作中提到】
: True awareness happens when the awareness sees itself.
M*n
58 楼
那些都是verified antibody, 不是专门为fish protein做的。
【在 a********k 的大作中提到】
: anaspec and GeneTex got hundreds antibodies for fish. Better than nothing.
【在 a********k 的大作中提到】
: anaspec and GeneTex got hundreds antibodies for fish. Better than nothing.
N*n
59 楼
You are welcome. I am not sure I can answer your questions, but I will try.
From your questions, I just realized that our discussion is a little too
advanced for you (don't get offended,:)), so i am going to back down to some
basics. I don't come to this board often and I don't know everyone's
background in Buddhism study. So, first, I want to ask you: do you do
meditation? A lot of things I am talking about above are based on meditation
experience. Without meditation, it is difficult to progress in the Buddhism
study. Second question, what kind of Buddhism text or books have you
studied?
First I should say thanks a lot for your detailed reply. :-)
Why does the awareness have the ability to know the room of mind is empty?
How to let the room of mind be empty?
>> Do you mean room or mind? Maybe you did not fully understand the example.
Maybe my example is not good example. :)
How could the mind be empty?
>>For begginers, it is difficult to detect. As you advance in meditation,
just let go of the thoughts, you can experience empty mind.
When the mind is empty, could the awareness still work?
>>Yes.
What is the physiological basis of our busy mind and the awareness?
>>Not sure what you mean. Just look at yourself, is your mind always
thinking about something?
Do they need similar mechanisms to run?
>>Again, not sure what you mean. Do you mean are they the function of the
brain cell? To me, I don't care. I just know from meditation experience and
buddhism study how mind and awareness work.
“无念”?
>>When mind is not thinking anything - no thought.
How to perform “无念”?
>> This is impossible for begginers. 念是心的功能,故无需刻意去造作无念.
How to judge the awareness is the true awareness?
>>For begginers, it is impossible to judge. It is also not neccessary. When
meditation, just be mindful of the meditation object.
Could you give a scientific definition or description of the awareness?
>>No, I don't have scientific definition of the awareness.
Waiting for more input from you. Thanks!
【在 J******s 的大作中提到】
: I was busy at work this morning and did not have time to type an example.
: Now I have a little bit of time and I will try to explain to you with an
: example. Suppose you are in a room. When you do not see anybody else in the
: room, you know you are the only one in the room. This same situation can be
: applied to awareness. Suppose this room has light - awareness is like the
: light. Let's say there is nothing in the room except light. The room is
: totally empty. Remember the awareness has the ability to know. Normally the
: awareness is always busy knowing all the things happening in the room. Now
: the room is totally empty. The awareness still try to know what's happening
: in the room - try to know. What awareness sees? Only emptiness and clarity (
From your questions, I just realized that our discussion is a little too
advanced for you (don't get offended,:)), so i am going to back down to some
basics. I don't come to this board often and I don't know everyone's
background in Buddhism study. So, first, I want to ask you: do you do
meditation? A lot of things I am talking about above are based on meditation
experience. Without meditation, it is difficult to progress in the Buddhism
study. Second question, what kind of Buddhism text or books have you
studied?
First I should say thanks a lot for your detailed reply. :-)
Why does the awareness have the ability to know the room of mind is empty?
How to let the room of mind be empty?
>> Do you mean room or mind? Maybe you did not fully understand the example.
Maybe my example is not good example. :)
How could the mind be empty?
>>For begginers, it is difficult to detect. As you advance in meditation,
just let go of the thoughts, you can experience empty mind.
When the mind is empty, could the awareness still work?
>>Yes.
What is the physiological basis of our busy mind and the awareness?
>>Not sure what you mean. Just look at yourself, is your mind always
thinking about something?
Do they need similar mechanisms to run?
>>Again, not sure what you mean. Do you mean are they the function of the
brain cell? To me, I don't care. I just know from meditation experience and
buddhism study how mind and awareness work.
“无念”?
>>When mind is not thinking anything - no thought.
How to perform “无念”?
>> This is impossible for begginers. 念是心的功能,故无需刻意去造作无念.
How to judge the awareness is the true awareness?
>>For begginers, it is impossible to judge. It is also not neccessary. When
meditation, just be mindful of the meditation object.
Could you give a scientific definition or description of the awareness?
>>No, I don't have scientific definition of the awareness.
Waiting for more input from you. Thanks!
【在 J******s 的大作中提到】
: I was busy at work this morning and did not have time to type an example.
: Now I have a little bit of time and I will try to explain to you with an
: example. Suppose you are in a room. When you do not see anybody else in the
: room, you know you are the only one in the room. This same situation can be
: applied to awareness. Suppose this room has light - awareness is like the
: light. Let's say there is nothing in the room except light. The room is
: totally empty. Remember the awareness has the ability to know. Normally the
: awareness is always busy knowing all the things happening in the room. Now
: the room is totally empty. The awareness still try to know what's happening
: in the room - try to know. What awareness sees? Only emptiness and clarity (
a*k
60 楼
credit肯定要给的，至于能给多大的话，我估计至少比rat KO的credit的零头多吧
【在 p*****m 的大作中提到】
: 嗯 我今天才看到个paper可以用在这个上面 哈 你觉得做出来这个会有多大用呢？对
: fish people来说，我一个外人没感觉。。
:
: 了。
【在 p*****m 的大作中提到】
: 嗯 我今天才看到个paper可以用在这个上面 哈 你觉得做出来这个会有多大用呢？对
: fish people来说，我一个外人没感觉。。
:
: 了。
N*n
61 楼
呵呵,有点意思. 看了你这帖,想起了洞山禅师的开悟偈:
切忌从他觅，迢迢与我殊。
我今独自往，处处得逢渠。
渠今正是我，我今不是渠。
应须恁么会，方可契如如。
这里渠就是指awareness.
【 在 bdbd (kakali) 的大作中提到: 】
awareness
切忌从他觅，迢迢与我殊。
我今独自往，处处得逢渠。
渠今正是我，我今不是渠。
应须恁么会，方可契如如。
这里渠就是指awareness.
【 在 bdbd (kakali) 的大作中提到: 】
awareness
a*k
62 楼
他们新做的，大部分都没有verify，如果你有兴趣verify，他们会免费给你的
【在 M*****n 的大作中提到】
: 那些都是verified antibody, 不是专门为fish protein做的。
【在 M*****n 的大作中提到】
: 那些都是verified antibody, 不是专门为fish protein做的。
J*s
63 楼
haha
Could I ask some similar questions to you? How many years did you practice
meditation and to which stage you are? Which kind of books did you read? It
is very nice to hear that you may practice meditation for quite long time.
However, what I can not understand in the discussion there are always such a
lot judgements unrelated to the question itself. All the great teachers I
met never answered my questions in this way.Convincing answers come from
decent logic, clear definitions, the peacefulness and confidence sensed by
the audience.
I was not offended. ha
I asked those questions which may seem quite simple, but actually not so
easy to answer. I asked them because I had some answers or my own opinions
in my mind.
The Buddhism I am learning is quite different with what you prefer. But I
still want to know how you think about those questions.
thanks anyway!
I have sth to do, and may discuss with you in details some days later.
I can give you my answers to those questions.
Best wishes!
.
some
meditation
Buddhism
【在 N*******n 的大作中提到】
: You are welcome. I am not sure I can answer your questions, but I will try.
: From your questions, I just realized that our discussion is a little too
: advanced for you (don't get offended,:)), so i am going to back down to some
: basics. I don't come to this board often and I don't know everyone's
: background in Buddhism study. So, first, I want to ask you: do you do
: meditation? A lot of things I am talking about above are based on meditation
: experience. Without meditation, it is difficult to progress in the Buddhism
: study. Second question, what kind of Buddhism text or books have you
: studied?
:
Could I ask some similar questions to you? How many years did you practice
meditation and to which stage you are? Which kind of books did you read? It
is very nice to hear that you may practice meditation for quite long time.
However, what I can not understand in the discussion there are always such a
lot judgements unrelated to the question itself. All the great teachers I
met never answered my questions in this way.Convincing answers come from
decent logic, clear definitions, the peacefulness and confidence sensed by
the audience.
I was not offended. ha
I asked those questions which may seem quite simple, but actually not so
easy to answer. I asked them because I had some answers or my own opinions
in my mind.
The Buddhism I am learning is quite different with what you prefer. But I
still want to know how you think about those questions.
thanks anyway!
I have sth to do, and may discuss with you in details some days later.
I can give you my answers to those questions.
Best wishes!
.
some
meditation
Buddhism
【在 N*******n 的大作中提到】
: You are welcome. I am not sure I can answer your questions, but I will try.
: From your questions, I just realized that our discussion is a little too
: advanced for you (don't get offended,:)), so i am going to back down to some
: basics. I don't come to this board often and I don't know everyone's
: background in Buddhism study. So, first, I want to ask you: do you do
: meditation? A lot of things I am talking about above are based on meditation
: experience. Without meditation, it is difficult to progress in the Buddhism
: study. Second question, what kind of Buddhism text or books have you
: studied?
:
p*m
64 楼
你这个冷水。。。鱼透明这个好处是替代不了的 而且人家体外受精 你们老鼠行么。。
呵呵
【在 s******y 的大作中提到】
: 偶知道，他现在不是想去做鱼么，所以我要给他浇点冷水。
: 鱼还有一个坏事，就是很多抗体都没有卖的。很多实验室都是索性自己做一个。
: 幸好据说互相之间不是很难借到。但是万一碰到不耐斯的人，就麻烦了。
呵呵
【在 s******y 的大作中提到】
: 偶知道，他现在不是想去做鱼么，所以我要给他浇点冷水。
: 鱼还有一个坏事，就是很多抗体都没有卖的。很多实验室都是索性自己做一个。
: 幸好据说互相之间不是很难借到。但是万一碰到不耐斯的人，就麻烦了。
N*n
65 楼
I appologize for a my last post to you. It was written in a hurry and with
bad attitude. Really sorry for that.
Anyway, thank you for helping me seeing myself and my grasping through the
discussion.
May I recommend a book to you: <
http://www.amazon.com/Joy-Living-Unlocking-Science-Happiness/dp
It's a great book good for both Buddhist and non-Buddhist.
Have a good day and best wishes.
It
a
【在 J******s 的大作中提到】
: haha
: Could I ask some similar questions to you? How many years did you practice
: meditation and to which stage you are? Which kind of books did you read? It
: is very nice to hear that you may practice meditation for quite long time.
: However, what I can not understand in the discussion there are always such a
: lot judgements unrelated to the question itself. All the great teachers I
: met never answered my questions in this way.Convincing answers come from
: decent logic, clear definitions, the peacefulness and confidence sensed by
: the audience.
: I was not offended. ha
bad attitude. Really sorry for that.
Anyway, thank you for helping me seeing myself and my grasping through the
discussion.
May I recommend a book to you: <
http://www.amazon.com/Joy-Living-Unlocking-Science-Happiness/dp
It's a great book good for both Buddhist and non-Buddhist.
Have a good day and best wishes.
It
a
【在 J******s 的大作中提到】
: haha
: Could I ask some similar questions to you? How many years did you practice
: meditation and to which stage you are? Which kind of books did you read? It
: is very nice to hear that you may practice meditation for quite long time.
: However, what I can not understand in the discussion there are always such a
: lot judgements unrelated to the question itself. All the great teachers I
: met never answered my questions in this way.Convincing answers come from
: decent logic, clear definitions, the peacefulness and confidence sensed by
: the audience.
: I was not offended. ha
p*m
66 楼
这么差？要说fish cKO应该还是挺有用的吧
【在 a********k 的大作中提到】
: credit肯定要给的，至于能给多大的话，我估计至少比rat KO的credit的零头多吧
【在 a********k 的大作中提到】
: credit肯定要给的，至于能给多大的话，我估计至少比rat KO的credit的零头多吧
b*d
67 楼
俺觉得你的问题在于你自己跟人讨论问题的时候，口气居高临下，不尊重他人。 all
the great teachers you see, I do not believe, they speak the way you do. 你
可以表达你的意见，但是与此同时，你必须明白别人的想法也是经过思考，值得尊重的
。 这只是给你的建议而已。
你说的什么不重要，而是你如何去说。 中国有句古话，一句话让人跳，一句话让人笑
。 I believe most people want others to 笑。。。。。。。。。
It
a
【在 J******s 的大作中提到】
: haha
: Could I ask some similar questions to you? How many years did you practice
: meditation and to which stage you are? Which kind of books did you read? It
: is very nice to hear that you may practice meditation for quite long time.
: However, what I can not understand in the discussion there are always such a
: lot judgements unrelated to the question itself. All the great teachers I
: met never answered my questions in this way.Convincing answers come from
: decent logic, clear definitions, the peacefulness and confidence sensed by
: the audience.
: I was not offended. ha
the great teachers you see, I do not believe, they speak the way you do. 你
可以表达你的意见，但是与此同时，你必须明白别人的想法也是经过思考，值得尊重的
。 这只是给你的建议而已。
你说的什么不重要，而是你如何去说。 中国有句古话，一句话让人跳，一句话让人笑
。 I believe most people want others to 笑。。。。。。。。。
It
a
【在 J******s 的大作中提到】
: haha
: Could I ask some similar questions to you? How many years did you practice
: meditation and to which stage you are? Which kind of books did you read? It
: is very nice to hear that you may practice meditation for quite long time.
: However, what I can not understand in the discussion there are always such a
: lot judgements unrelated to the question itself. All the great teachers I
: met never answered my questions in this way.Convincing answers come from
: decent logic, clear definitions, the peacefulness and confidence sensed by
: the audience.
: I was not offended. ha
O*e
68 楼
以后我如果有闲工夫就搞搞这个老鼠的体外受精，呵呵
【在 p*****m 的大作中提到】
: 你这个冷水。。。鱼透明这个好处是替代不了的 而且人家体外受精 你们老鼠行么。。
: 呵呵
【在 p*****m 的大作中提到】
: 你这个冷水。。。鱼透明这个好处是替代不了的 而且人家体外受精 你们老鼠行么。。
: 呵呵
N*n
69 楼
Jean的这一贴是因为我的回贴态度不好才引起的,所以是我的不对.我已经给她道歉了,
所以大家就不用再互相关爱了.:)
【在 b**d 的大作中提到】
: 俺觉得你的问题在于你自己跟人讨论问题的时候，口气居高临下，不尊重他人。 all
: the great teachers you see, I do not believe, they speak the way you do. 你
: 可以表达你的意见，但是与此同时，你必须明白别人的想法也是经过思考，值得尊重的
: 。 这只是给你的建议而已。
: 你说的什么不重要，而是你如何去说。 中国有句古话，一句话让人跳，一句话让人笑
: 。 I believe most people want others to 笑。。。。。。。。。
:
: It
: a
所以大家就不用再互相关爱了.:)
【在 b**d 的大作中提到】
: 俺觉得你的问题在于你自己跟人讨论问题的时候，口气居高临下，不尊重他人。 all
: the great teachers you see, I do not believe, they speak the way you do. 你
: 可以表达你的意见，但是与此同时，你必须明白别人的想法也是经过思考，值得尊重的
: 。 这只是给你的建议而已。
: 你说的什么不重要，而是你如何去说。 中国有句古话，一句话让人跳，一句话让人笑
: 。 I believe most people want others to 笑。。。。。。。。。
:
: It
: a
a*k
70 楼
个人意见。。。从悲观的角度出发而已
【在 p*****m 的大作中提到】
: 这么差？要说fish cKO应该还是挺有用的吧
【在 p*****m 的大作中提到】
: 这么差？要说fish cKO应该还是挺有用的吧
b*d
71 楼
俺觉得她挺不错的，很爱学，讲东西也蛮有道理的，但是就因为说话口气太直，让人误
会，比较可惜，所以才说上面这些话的。 没有要攻击她的意思。估计俺说话也太直啦
。 ：）
【在 N*******n 的大作中提到】
: Jean的这一贴是因为我的回贴态度不好才引起的,所以是我的不对.我已经给她道歉了,
: 所以大家就不用再互相关爱了.:)
会，比较可惜，所以才说上面这些话的。 没有要攻击她的意思。估计俺说话也太直啦
。 ：）
【在 N*******n 的大作中提到】
: Jean的这一贴是因为我的回贴态度不好才引起的,所以是我的不对.我已经给她道歉了,
: 所以大家就不用再互相关爱了.:)
s*y
72 楼
关键是做出来之后又不知道和人的系统有没有差别。
弄不好又白做了。白白发展一个技术作出来的东西都不知道有什么意义。
否则大家干吗拼死拼活的要上老鼠和大鼠？图的就是这个哺乳动物和人比较接近。
【在 p*****m 的大作中提到】
: 这么差？要说fish cKO应该还是挺有用的吧
弄不好又白做了。白白发展一个技术作出来的东西都不知道有什么意义。
否则大家干吗拼死拼活的要上老鼠和大鼠？图的就是这个哺乳动物和人比较接近。
【在 p*****m 的大作中提到】
: 这么差？要说fish cKO应该还是挺有用的吧
J*s
73 楼
现在国内,翻过来的,就不多敲字了.谢谢推荐,我会认真读的,哈
:-)
其实我对你态度没有太大质疑了,不过我个人就比较喜欢就问题讨论问题
当然我讨论问题方式态度也有待改善,哈
and
【在 N*******n 的大作中提到】
: I appologize for a my last post to you. It was written in a hurry and with
: bad attitude. Really sorry for that.
: Anyway, thank you for helping me seeing myself and my grasping through the
: discussion.
: May I recommend a book to you: <
: http://www.amazon.com/Joy-Living-Unlocking-Science-Happiness/dp
: It's a great book good for both Buddhist and non-Buddhist.
: Have a good day and best wishes.
:
:-)
其实我对你态度没有太大质疑了,不过我个人就比较喜欢就问题讨论问题
当然我讨论问题方式态度也有待改善,哈
and
【在 N*******n 的大作中提到】
: I appologize for a my last post to you. It was written in a hurry and with
: bad attitude. Really sorry for that.
: Anyway, thank you for helping me seeing myself and my grasping through the
: discussion.
: May I recommend a book to you: <
: http://www.amazon.com/Joy-Living-Unlocking-Science-Happiness/dp
: It's a great book good for both Buddhist and non-Buddhist.
: Have a good day and best wishes.
:
p*m
74 楼
要我说这个是urban legend，rodent比human还是差得远
而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
头都不如
【在 s******y 的大作中提到】
: 关键是做出来之后又不知道和人的系统有没有差别。
: 弄不好又白做了。白白发展一个技术作出来的东西都不知道有什么意义。
: 否则大家干吗拼死拼活的要上老鼠和大鼠？图的就是这个哺乳动物和人比较接近。
而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
头都不如
【在 s******y 的大作中提到】
: 关键是做出来之后又不知道和人的系统有没有差别。
: 弄不好又白做了。白白发展一个技术作出来的东西都不知道有什么意义。
: 否则大家干吗拼死拼活的要上老鼠和大鼠？图的就是这个哺乳动物和人比较接近。
J*s
75 楼
多谢指出缺点哈
我没居高临下的意思,不过我大概知道自己的问题了
我说的是好老师从不拿资历身份来讨论问题,当然老师大多数情况是合适不合适的区别
我给别人意见credit了
:-)
我觉得说什么和怎么说一样重要,你说的对方式确实很重要,但是取决于我们说话的目的
至于是让人跳还是笑,我理解的是让听众觉得舒服更好
thanks!
【在 b**d 的大作中提到】
: 俺觉得你的问题在于你自己跟人讨论问题的时候，口气居高临下，不尊重他人。 all
: the great teachers you see, I do not believe, they speak the way you do. 你
: 可以表达你的意见，但是与此同时，你必须明白别人的想法也是经过思考，值得尊重的
: 。 这只是给你的建议而已。
: 你说的什么不重要，而是你如何去说。 中国有句古话，一句话让人跳，一句话让人笑
: 。 I believe most people want others to 笑。。。。。。。。。
:
: It
: a
我没居高临下的意思,不过我大概知道自己的问题了
我说的是好老师从不拿资历身份来讨论问题,当然老师大多数情况是合适不合适的区别
我给别人意见credit了
:-)
我觉得说什么和怎么说一样重要,你说的对方式确实很重要,但是取决于我们说话的目的
至于是让人跳还是笑,我理解的是让听众觉得舒服更好
thanks!
【在 b**d 的大作中提到】
: 俺觉得你的问题在于你自己跟人讨论问题的时候，口气居高临下，不尊重他人。 all
: the great teachers you see, I do not believe, they speak the way you do. 你
: 可以表达你的意见，但是与此同时，你必须明白别人的想法也是经过思考，值得尊重的
: 。 这只是给你的建议而已。
: 你说的什么不重要，而是你如何去说。 中国有句古话，一句话让人跳，一句话让人笑
: 。 I believe most people want others to 笑。。。。。。。。。
:
: It
: a
p*m
76 楼
哦。。
【在 a********k 的大作中提到】
: 个人意见。。。从悲观的角度出发而已
【在 a********k 的大作中提到】
: 个人意见。。。从悲观的角度出发而已
J*s
77 楼
never mind:-)
【在 b**d 的大作中提到】
: 俺觉得她挺不错的，很爱学，讲东西也蛮有道理的，但是就因为说话口气太直，让人误
: 会，比较可惜，所以才说上面这些话的。 没有要攻击她的意思。估计俺说话也太直啦
: 。 ：）
【在 b**d 的大作中提到】
: 俺觉得她挺不错的，很爱学，讲东西也蛮有道理的，但是就因为说话口气太直，让人误
: 会，比较可惜，所以才说上面这些话的。 没有要攻击她的意思。估计俺说话也太直啦
: 。 ：）
a*k
78 楼
但是你就觉得鱼比rodent有更多贡献？线虫和果蝇的贡献咱暂且不提，鱼到现在基本可
以说是零贡献。。。有了cKO以后有没有零头还是个问题
的零
【在 p*****m 的大作中提到】
: 要我说这个是urban legend，rodent比human还是差得远
: 而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
: 头都不如
以说是零贡献。。。有了cKO以后有没有零头还是个问题
的零
【在 p*****m 的大作中提到】
: 要我说这个是urban legend，rodent比human还是差得远
: 而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
: 头都不如
N*n
79 楼
我给你推荐的那本书，国内能买到中文版的，书名叫《世界上最快乐的人》。非常好的
讲述了禅修与生活的融合。如果你读了，有问题可以贴到版上来，我们大家一起学习讨
论。
【在 J******s 的大作中提到】
: 现在国内,翻过来的,就不多敲字了.谢谢推荐,我会认真读的,哈
: :-)
: 其实我对你态度没有太大质疑了,不过我个人就比较喜欢就问题讨论问题
: 当然我讨论问题方式态度也有待改善,哈
:
: and
讲述了禅修与生活的融合。如果你读了，有问题可以贴到版上来，我们大家一起学习讨
论。
【在 J******s 的大作中提到】
: 现在国内,翻过来的,就不多敲字了.谢谢推荐,我会认真读的,哈
: :-)
: 其实我对你态度没有太大质疑了,不过我个人就比较喜欢就问题讨论问题
: 当然我讨论问题方式态度也有待改善,哈
:
: and
s*r
80 楼
ZFN做knock-in的话大片段像2kb以上就不太好插入了，而且效率也低，做老鼠的话打那
么多embryo，还要都生出来做genotyping,都是钱啊... 鱼的我不懂，可能便宜很多吧
么多embryo，还要都生出来做genotyping,都是钱啊... 鱼的我不懂，可能便宜很多吧
s*y
81 楼
rodent 好歹是个哺乳动物啊。
鱼啊什么的和人也差的太远了
的零
【在 p*****m 的大作中提到】
: 要我说这个是urban legend，rodent比human还是差得远
: 而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
: 头都不如
鱼啊什么的和人也差的太远了
的零
【在 p*****m 的大作中提到】
: 要我说这个是urban legend，rodent比human还是差得远
: 而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
: 头都不如
A*y
82 楼
Sigma will do it for ~25k...any gene anywhere as far as I understood.
【在 M*****n 的大作中提到】
: knock-in is a plus, 但是knock-out 最重要。
: 现在ZFN的效率太低了， 最多20%。你如果能弄一个比较efficiet的ZFN,比如80%, 那么
: knock-in有没有也无所谓。
:
: 要的
【在 M*****n 的大作中提到】
: knock-in is a plus, 但是knock-out 最重要。
: 现在ZFN的效率太低了， 最多20%。你如果能弄一个比较efficiet的ZFN,比如80%, 那么
: knock-in有没有也无所谓。
:
: 要的
p*m
83 楼
哦 这个是自然 fish的贡献就是0...所以我才不sure这个有多有意义
话说回来，不管有没有意义 只要fish community觉得有用给我个job就行啊 也不用非
要想得那么长远不是 呵呵
【在 a********k 的大作中提到】
: 但是你就觉得鱼比rodent有更多贡献？线虫和果蝇的贡献咱暂且不提，鱼到现在基本可
: 以说是零贡献。。。有了cKO以后有没有零头还是个问题
:
: 的零
话说回来，不管有没有意义 只要fish community觉得有用给我个job就行啊 也不用非
要想得那么长远不是 呵呵
【在 a********k 的大作中提到】
: 但是你就觉得鱼比rodent有更多贡献？线虫和果蝇的贡献咱暂且不提，鱼到现在基本可
: 以说是零贡献。。。有了cKO以后有没有零头还是个问题
:
: 的零
p*m
84 楼
用ZFN做直接knockin是不行的（也就老鼠还行，人细胞和fish的HM比率都非常低）
【在 s********r 的大作中提到】
: ZFN做knock-in的话大片段像2kb以上就不太好插入了，而且效率也低，做老鼠的话打那
: 么多embryo，还要都生出来做genotyping,都是钱啊... 鱼的我不懂，可能便宜很多吧
【在 s********r 的大作中提到】
: ZFN做knock-in的话大片段像2kb以上就不太好插入了，而且效率也低，做老鼠的话打那
: 么多embryo，还要都生出来做genotyping,都是钱啊... 鱼的我不懂，可能便宜很多吧
a*k
85 楼
显然会有人给job啊，而且不是少数
搞tool的几个fish lab都混得不错啊，搞出Tol2的日本人就不说了
发展Tol2的chien在fish community里面就很有地位了
其他搞Tool的人，做MO的Ekker，搞出virus insertion的hopkins都很有地位
【在 p*****m 的大作中提到】
: 哦 这个是自然 fish的贡献就是0...所以我才不sure这个有多有意义
: 话说回来，不管有没有意义 只要fish community觉得有用给我个job就行啊 也不用非
: 要想得那么长远不是 呵呵
搞tool的几个fish lab都混得不错啊，搞出Tol2的日本人就不说了
发展Tol2的chien在fish community里面就很有地位了
其他搞Tool的人，做MO的Ekker，搞出virus insertion的hopkins都很有地位
【在 p*****m 的大作中提到】
: 哦 这个是自然 fish的贡献就是0...所以我才不sure这个有多有意义
: 话说回来，不管有没有意义 只要fish community觉得有用给我个job就行啊 也不用非
: 要想得那么长远不是 呵呵
p*m
86 楼
then it is good enough, lol
啥时候msn找你聊哈
【在 a********k 的大作中提到】
: 显然会有人给job啊，而且不是少数
: 搞tool的几个fish lab都混得不错啊，搞出Tol2的日本人就不说了
: 发展Tol2的chien在fish community里面就很有地位了
: 其他搞Tool的人，做MO的Ekker，搞出virus insertion的hopkins都很有地位
啥时候msn找你聊哈
【在 a********k 的大作中提到】
: 显然会有人给job啊，而且不是少数
: 搞tool的几个fish lab都混得不错啊，搞出Tol2的日本人就不说了
: 发展Tol2的chien在fish community里面就很有地位了
: 其他搞Tool的人，做MO的Ekker，搞出virus insertion的hopkins都很有地位
M*n
87 楼
Chi-Bin Chien, Steve Ekker, Kawakami不光是搞了tool.
他们搞这些tool之前早就有地位了。
Nancy Hopkins也就是资格老一点，其实没作出什么大的东西。那个insertion的idea也
很普通。
【在 a********k 的大作中提到】
: 显然会有人给job啊，而且不是少数
: 搞tool的几个fish lab都混得不错啊，搞出Tol2的日本人就不说了
: 发展Tol2的chien在fish community里面就很有地位了
: 其他搞Tool的人，做MO的Ekker，搞出virus insertion的hopkins都很有地位
他们搞这些tool之前早就有地位了。
Nancy Hopkins也就是资格老一点，其实没作出什么大的东西。那个insertion的idea也
很普通。
【在 a********k 的大作中提到】
: 显然会有人给job啊，而且不是少数
: 搞tool的几个fish lab都混得不错啊，搞出Tol2的日本人就不说了
: 发展Tol2的chien在fish community里面就很有地位了
: 其他搞Tool的人，做MO的Ekker，搞出virus insertion的hopkins都很有地位
M*n
88 楼
Sigma will validate the ZFN in vitro, you have to test it in fish by
yourself.
it is too risky and too expensive.
【在 A******y 的大作中提到】
: Sigma will do it for ~25k...any gene anywhere as far as I understood.
yourself.
it is too risky and too expensive.
【在 A******y 的大作中提到】
: Sigma will do it for ~25k...any gene anywhere as far as I understood.
a*k
89 楼
请教一下Chien, Ekker, Kawakami, Hopkins比Tool明显高一个档次的贡献。
Hopkins也算是纯做zebrafish唯一的院士了吧，其实整个领域就没作出啥东西。。。
【在 M*****n 的大作中提到】
: Chi-Bin Chien, Steve Ekker, Kawakami不光是搞了tool.
: 他们搞这些tool之前早就有地位了。
: Nancy Hopkins也就是资格老一点，其实没作出什么大的东西。那个insertion的idea也
: 很普通。
Hopkins也算是纯做zebrafish唯一的院士了吧，其实整个领域就没作出啥东西。。。
【在 M*****n 的大作中提到】
: Chi-Bin Chien, Steve Ekker, Kawakami不光是搞了tool.
: 他们搞这些tool之前早就有地位了。
: Nancy Hopkins也就是资格老一点，其实没作出什么大的东西。那个insertion的idea也
: 很普通。
f*u
90 楼
如果你先用ZFN来KI一个attP，然后再把你要的片断钉进去怎么样
【在 p*****m 的大作中提到】
: 用ZFN做直接knockin是不行的（也就老鼠还行，人细胞和fish的HM比率都非常低）
【在 p*****m 的大作中提到】
: 用ZFN做直接knockin是不行的（也就老鼠还行，人细胞和fish的HM比率都非常低）
f*u
91 楼
我看了几个帖子有点晕了，ZFN的目的不就是增加HR的效率吗？
难道在鱼里面，ZFN有什么其它的用处？
【在 M*****n 的大作中提到】
: ZFN不如homologous rec好。
: 有很多limitation.
: 你再读paper哦。
难道在鱼里面，ZFN有什么其它的用处？
【在 M*****n 的大作中提到】
: ZFN不如homologous rec好。
: 有很多limitation.
: 你再读paper哦。
F*K
92 楼
给科普一下Ekker的贡献把，看来我是有眼不识泰山了。。
【在 M*****n 的大作中提到】
: Chi-Bin Chien, Steve Ekker, Kawakami不光是搞了tool.
: 他们搞这些tool之前早就有地位了。
: Nancy Hopkins也就是资格老一点，其实没作出什么大的东西。那个insertion的idea也
: 很普通。
【在 M*****n 的大作中提到】
: Chi-Bin Chien, Steve Ekker, Kawakami不光是搞了tool.
: 他们搞这些tool之前早就有地位了。
: Nancy Hopkins也就是资格老一点，其实没作出什么大的东西。那个insertion的idea也
: 很普通。
f*u
93 楼
如果你下决心做鱼了，我看不如把MARCM给移植到鱼里面，这样长期的贡献应该不小。
【在 p*****m 的大作中提到】
: 哦 这个是自然 fish的贡献就是0...所以我才不sure这个有多有意义
: 话说回来，不管有没有意义 只要fish community觉得有用给我个job就行啊 也不用非
: 要想得那么长远不是 呵呵
【在 p*****m 的大作中提到】
: 哦 这个是自然 fish的贡献就是0...所以我才不sure这个有多有意义
: 话说回来，不管有没有意义 只要fish community觉得有用给我个job就行啊 也不用非
: 要想得那么长远不是 呵呵
a*k
94 楼
MARCM不是那么容易移植的。。。没有cKO移植了贡献有限
而且鱼的工作绝大部分在幼鱼里面做，要trace cell linage基本用transient
expression或者transplantation了。MARCM的优势比起在老鼠里面小无数倍。
【在 f**u 的大作中提到】
: 如果你下决心做鱼了，我看不如把MARCM给移植到鱼里面，这样长期的贡献应该不小。
而且鱼的工作绝大部分在幼鱼里面做，要trace cell linage基本用transient
expression或者transplantation了。MARCM的优势比起在老鼠里面小无数倍。
【在 f**u 的大作中提到】
: 如果你下决心做鱼了，我看不如把MARCM给移植到鱼里面，这样长期的贡献应该不小。
i*g
95 楼
问一个问题，
我现在一个相关project是 zfn， target 就不说了，虽然说了也没有什么。
问题是经理是个 adeno viral控，什么都往那里面装
这个zfn对adeno packing有多少影响？ 是否包装困难？（当然，我知道文献有成功报
导，不过，我历来是非常negative,）
我现在一个相关project是 zfn， target 就不说了，虽然说了也没有什么。
问题是经理是个 adeno viral控，什么都往那里面装
这个zfn对adeno packing有多少影响？ 是否包装困难？（当然，我知道文献有成功报
导，不过，我历来是非常negative,）
g*5
96 楼
haha
摆摊卖鱼了
【在 s******y 的大作中提到】
: 鱼的好处就是身体透明，而且发育过程都在体外吧
: 要是什么时候做老鼠的搞出了人造老鼠子宫，你们做鱼的就可以去摆摊卖鱼了。
:
: model
: transpare
: rep
: to
: 很多
: serious的
: 大好
摆摊卖鱼了
【在 s******y 的大作中提到】
: 鱼的好处就是身体透明，而且发育过程都在体外吧
: 要是什么时候做老鼠的搞出了人造老鼠子宫，你们做鱼的就可以去摆摊卖鱼了。
:
: model
: transpare
: rep
: to
: 很多
: serious的
: 大好
O*e
97 楼
你要这样说，大肠杆菌的贡献可比线虫和果蝇的还要大。
这都是历史发展的不同阶段；可要是倒过来走，似乎就有些说不通了。
的零
【在 p*****m 的大作中提到】
: 要我说这个是urban legend，rodent比human还是差得远
: 而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
: 头都不如
这都是历史发展的不同阶段；可要是倒过来走，似乎就有些说不通了。
的零
【在 p*****m 的大作中提到】
: 要我说这个是urban legend，rodent比human还是差得远
: 而且 从我一个fly person的眼光看来，rodent model对生物学的贡献比线虫和果蝇的零
: 头都不如
A*A
98 楼
具我所知，几个fish labs正在联合用ZFN搞大规模的knock out resources。ZFN以及相
关技术最近会有比较大的进展。大家如果感兴趣，可以关注一下最近的TALEN技术。在
最近1-2年内，估计任何试验室都可以自己合成reagent去target任何基因。cost不会比
买限制性内切酶高太多。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
关技术最近会有比较大的进展。大家如果感兴趣，可以关注一下最近的TALEN技术。在
最近1-2年内，估计任何试验室都可以自己合成reagent去target任何基因。cost不会比
买限制性内切酶高太多。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
l*n
99 楼
在fish lab钓鱼是不是很有乐趣？
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
p*m
100 楼
bingo!
【在 f**u 的大作中提到】
: 如果你先用ZFN来KI一个attP，然后再把你要的片断钉进去怎么样
【在 f**u 的大作中提到】
: 如果你先用ZFN来KI一个attP，然后再把你要的片断钉进去怎么样
p*m
101 楼
ZFN在鱼里目前就是用来造成NHEJ来形成mutation的 你说这个增加HR效率的事情 目前
好像还停留在我脑子里 当然也许已经有人开做了:)
【在 f**u 的大作中提到】
: 我看了几个帖子有点晕了，ZFN的目的不就是增加HR的效率吗？
: 难道在鱼里面，ZFN有什么其它的用处？
好像还停留在我脑子里 当然也许已经有人开做了:)
【在 f**u 的大作中提到】
: 我看了几个帖子有点晕了，ZFN的目的不就是增加HR的效率吗？
: 难道在鱼里面，ZFN有什么其它的用处？
p*m
102 楼
MARCM困难 但是用基于cre的方法应该还好吧（就像老鼠的MADM） 问题就像你说的 这
个东西感觉不大有用 transient expression本来就已经mosaic了反正。就像marcm在果
蝇里用的很广泛 但是madm就没有什么人care，因为老鼠大家随便用virus搞mosaic
【在 a********k 的大作中提到】
: MARCM不是那么容易移植的。。。没有cKO移植了贡献有限
: 而且鱼的工作绝大部分在幼鱼里面做，要trace cell linage基本用transient
: expression或者transplantation了。MARCM的优势比起在老鼠里面小无数倍。
个东西感觉不大有用 transient expression本来就已经mosaic了反正。就像marcm在果
蝇里用的很广泛 但是madm就没有什么人care，因为老鼠大家随便用virus搞mosaic
【在 a********k 的大作中提到】
: MARCM不是那么容易移植的。。。没有cKO移植了贡献有限
: 而且鱼的工作绝大部分在幼鱼里面做，要trace cell linage基本用transient
: expression或者transplantation了。MARCM的优势比起在老鼠里面小无数倍。
p*m
103 楼
倒过来走谈不上吧 线虫上一个贡献的nobel prize才不过是几年前的事情(08年martin
chalfie) 再上一个也没几年（06 Fire&Mello）。这个和E Coli是完全不一样的。。再
说近的，过去10年对小RNA的理解，从miRNA到piRNA，都是线虫+果蝇里来的；Janelia更
是卯足了劲要把fly的事情搞清楚。这两个model system的用处还远没有开发完呢
【在 O******e 的大作中提到】
: 你要这样说，大肠杆菌的贡献可比线虫和果蝇的还要大。
: 这都是历史发展的不同阶段；可要是倒过来走，似乎就有些说不通了。
:
: 的零
chalfie) 再上一个也没几年（06 Fire&Mello）。这个和E Coli是完全不一样的。。再
说近的，过去10年对小RNA的理解，从miRNA到piRNA，都是线虫+果蝇里来的；Janelia更
是卯足了劲要把fly的事情搞清楚。这两个model system的用处还远没有开发完呢
【在 O******e 的大作中提到】
: 你要这样说，大肠杆菌的贡献可比线虫和果蝇的还要大。
: 这都是历史发展的不同阶段；可要是倒过来走，似乎就有些说不通了。
:
: 的零
p*m
104 楼
ZFN+TALEN做KO还是可以的 不过就像Morphin说的，光KO感觉并没啥太大意思了。 要说
fish mutants也大把大把的了 现在缺的并不是一个简单的KO工具感觉。有KI和cKO也许
能好一点
TALEN本来我想在鱼里做一下来着 就当练手 结果还没谈好合作就被发掉了(上周scienc
e, barbara Meyer在线虫里做了) 我就没兴趣在fish里再proof of principle一把了。
。而且也听说有别的fish lab做
【在 A****A 的大作中提到】
: 具我所知，几个fish labs正在联合用ZFN搞大规模的knock out resources。ZFN以及相
: 关技术最近会有比较大的进展。大家如果感兴趣，可以关注一下最近的TALEN技术。在
: 最近1-2年内，估计任何试验室都可以自己合成reagent去target任何基因。cost不会比
: 买限制性内切酶高太多。
:
: model
: transpare
: rep
: to
: 很多
fish mutants也大把大把的了 现在缺的并不是一个简单的KO工具感觉。有KI和cKO也许
能好一点
TALEN本来我想在鱼里做一下来着 就当练手 结果还没谈好合作就被发掉了(上周scienc
e, barbara Meyer在线虫里做了) 我就没兴趣在fish里再proof of principle一把了。
。而且也听说有别的fish lab做
【在 A****A 的大作中提到】
: 具我所知，几个fish labs正在联合用ZFN搞大规模的knock out resources。ZFN以及相
: 关技术最近会有比较大的进展。大家如果感兴趣，可以关注一下最近的TALEN技术。在
: 最近1-2年内，估计任何试验室都可以自己合成reagent去target任何基因。cost不会比
: 买限制性内切酶高太多。
:
: model
: transpare
: rep
: to
: 很多
y*8
105 楼
Chemical screening is so powerful in fish and probably the only thing worth
pursuing if you want a easy-pass for your career. I have seen lots of young
fish scientists miserably failed by just doing genetics.
pursuing if you want a easy-pass for your career. I have seen lots of young
fish scientists miserably failed by just doing genetics.
p*m
106 楼
我差不多现在就是这个想法哈
worth
young
【在 y******8 的大作中提到】
: Chemical screening is so powerful in fish and probably the only thing worth
: pursuing if you want a easy-pass for your career. I have seen lots of young
: fish scientists miserably failed by just doing genetics.
worth
young
【在 y******8 的大作中提到】
: Chemical screening is so powerful in fish and probably the only thing worth
: pursuing if you want a easy-pass for your career. I have seen lots of young
: fish scientists miserably failed by just doing genetics.
X*n
107 楼
感觉fish这个model如果真会有贡献的话，可能最主要会在chemical genomics方面，在
血液病方面可能也有些优势。其它方面没觉得有很大优势。一些大动物模型会慢慢发展
起来，觉得国内在这方面有很大优势。
血液病方面可能也有些优势。其它方面没觉得有很大优势。一些大动物模型会慢慢发展
起来，觉得国内在这方面有很大优势。
a*k
108 楼
血液也很难讲。。。不是做血液的，但是不看好。
光从现在的结果来看，鱼的血液发育和哺乳类差别很大了
之所以现在做的人那么多是因为primitive hematopoeisis在老鼠里面压根没法做，鱼
胚胎用来做卵黄囊造血是再好不过的model了。但是血液病大多数还是骨髓干细胞的问
题，感觉用鱼来做这个也就是个忽悠。不知道做血液的怎么看？
【在 X******n 的大作中提到】
: 感觉fish这个model如果真会有贡献的话，可能最主要会在chemical genomics方面，在
: 血液病方面可能也有些优势。其它方面没觉得有很大优势。一些大动物模型会慢慢发展
: 起来，觉得国内在这方面有很大优势。
光从现在的结果来看，鱼的血液发育和哺乳类差别很大了
之所以现在做的人那么多是因为primitive hematopoeisis在老鼠里面压根没法做，鱼
胚胎用来做卵黄囊造血是再好不过的model了。但是血液病大多数还是骨髓干细胞的问
题，感觉用鱼来做这个也就是个忽悠。不知道做血液的怎么看？
【在 X******n 的大作中提到】
: 感觉fish这个model如果真会有贡献的话，可能最主要会在chemical genomics方面，在
: 血液病方面可能也有些优势。其它方面没觉得有很大优势。一些大动物模型会慢慢发展
: 起来，觉得国内在这方面有很大优势。
y*8
109 楼
No only for hematopoietic system. Basically, you can screen for any
phenotype, but use soluble molecules instead of genetic methods. By this way
, you would directly get drug candidates, not just drug target candidates.
The transparency of fish embryo is overstated. If you want deal with any
late-onset disease, the adult fish is not very transparent.
【在 X******n 的大作中提到】
: 感觉fish这个model如果真会有贡献的话，可能最主要会在chemical genomics方面，在
: 血液病方面可能也有些优势。其它方面没觉得有很大优势。一些大动物模型会慢慢发展
: 起来，觉得国内在这方面有很大优势。
phenotype, but use soluble molecules instead of genetic methods. By this way
, you would directly get drug candidates, not just drug target candidates.
The transparency of fish embryo is overstated. If you want deal with any
late-onset disease, the adult fish is not very transparent.
【在 X******n 的大作中提到】
: 感觉fish这个model如果真会有贡献的话，可能最主要会在chemical genomics方面，在
: 血液病方面可能也有些优势。其它方面没觉得有很大优势。一些大动物模型会慢慢发展
: 起来，觉得国内在这方面有很大优势。
X*n
110 楼
you would directly get drug candidates, not just drug target candidates.
展开谈谈？
展开谈谈？
p*m
111 楼
这个只能说是科幻吧 不觉得fish small molecule screen能直接拿到drug candidate
way
【在 y******8 的大作中提到】
: No only for hematopoietic system. Basically, you can screen for any
: phenotype, but use soluble molecules instead of genetic methods. By this way
: , you would directly get drug candidates, not just drug target candidates.
: The transparency of fish embryo is overstated. If you want deal with any
: late-onset disease, the adult fish is not very transparent.
way
【在 y******8 的大作中提到】
: No only for hematopoietic system. Basically, you can screen for any
: phenotype, but use soluble molecules instead of genetic methods. By this way
: , you would directly get drug candidates, not just drug target candidates.
: The transparency of fish embryo is overstated. If you want deal with any
: late-onset disease, the adult fish is not very transparent.
A*A
112 楼
利用ZFN等reagents做Knock in应该是一些lab目前的奋斗方向。ZFN可以产生sequence
specific DNA double strand break. DNA双链断裂可以有效的提高homologous
recombination的频率，从而达到Knock in的目的。目前，在fish里面实现高效的HR还
很困难，即使在有ZFN的帮助下。我不是做fish的，主要做DNA repair pathway. 我觉
得可以通过一些genetic或chemical的手段抑制fish的NHEJ pathway，来增加HR的频率。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
specific DNA double strand break. DNA双链断裂可以有效的提高homologous
recombination的频率，从而达到Knock in的目的。目前，在fish里面实现高效的HR还
很困难，即使在有ZFN的帮助下。我不是做fish的，主要做DNA repair pathway. 我觉
得可以通过一些genetic或chemical的手段抑制fish的NHEJ pathway，来增加HR的频率。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
y*8
113 楼
You could modify these small molecules into real drugs. If you have a good
synthetic chemistry lab to cooperate with, why not.
candidate
【在 p*****m 的大作中提到】
: 这个只能说是科幻吧 不觉得fish small molecule screen能直接拿到drug candidate
:
: way
synthetic chemistry lab to cooperate with, why not.
candidate
【在 p*****m 的大作中提到】
: 这个只能说是科幻吧 不觉得fish small molecule screen能直接拿到drug candidate
:
: way
p*m
114 楼
嗯 这几天刚有paper出来show这个的。请问一下有什么具体思路来一直NHEJ,然后这个
为什么能促进HR呢？我对这些分子生物学东东一窍不通 要是有相关文献就更好了哈 多
谢多谢
sequence
率。
【在 A****A 的大作中提到】
: 利用ZFN等reagents做Knock in应该是一些lab目前的奋斗方向。ZFN可以产生sequence
: specific DNA double strand break. DNA双链断裂可以有效的提高homologous
: recombination的频率，从而达到Knock in的目的。目前，在fish里面实现高效的HR还
: 很困难，即使在有ZFN的帮助下。我不是做fish的，主要做DNA repair pathway. 我觉
: 得可以通过一些genetic或chemical的手段抑制fish的NHEJ pathway，来增加HR的频率。
:
: model
: transpare
: rep
: to
为什么能促进HR呢？我对这些分子生物学东东一窍不通 要是有相关文献就更好了哈 多
谢多谢
sequence
率。
【在 A****A 的大作中提到】
: 利用ZFN等reagents做Knock in应该是一些lab目前的奋斗方向。ZFN可以产生sequence
: specific DNA double strand break. DNA双链断裂可以有效的提高homologous
: recombination的频率，从而达到Knock in的目的。目前，在fish里面实现高效的HR还
: 很困难，即使在有ZFN的帮助下。我不是做fish的，主要做DNA repair pathway. 我觉
: 得可以通过一些genetic或chemical的手段抑制fish的NHEJ pathway，来增加HR的频率。
:
: model
: transpare
: rep
: to
A*y
115 楼
Drug screen is possible in zebrafish, I have one of this in my collaborator'
s lab. 96 wells zebrafish behavior monitoring chamber.
http://www.noldus.com/animal-behavior-research/products/daniovi
s lab. 96 wells zebrafish behavior monitoring chamber.
http://www.noldus.com/animal-behavior-research/products/daniovi
X*n
116 楼
从drug screen 到Drug candidates，还有很长的路要走。
collaborator'
【在 A******y 的大作中提到】
: Drug screen is possible in zebrafish, I have one of this in my collaborator'
: s lab. 96 wells zebrafish behavior monitoring chamber.
: http://www.noldus.com/animal-behavior-research/products/daniovi
collaborator'
【在 A******y 的大作中提到】
: Drug screen is possible in zebrafish, I have one of this in my collaborator'
: s lab. 96 wells zebrafish behavior monitoring chamber.
: http://www.noldus.com/animal-behavior-research/products/daniovi
A*y
117 楼
True, but it is good enough to publish paper :)
【在 X******n 的大作中提到】
: 从drug screen 到Drug candidates，还有很长的路要走。
:
: collaborator'
【在 X******n 的大作中提到】
: 从drug screen 到Drug candidates，还有很长的路要走。
:
: collaborator'
G*m
118 楼
插个loxP,attP之类的就很好用了。。。
【在 s********r 的大作中提到】
: ZFN做knock-in的话大片段像2kb以上就不太好插入了，而且效率也低，做老鼠的话打那
: 么多embryo，还要都生出来做genotyping,都是钱啊... 鱼的我不懂，可能便宜很多吧
【在 s********r 的大作中提到】
: ZFN做knock-in的话大片段像2kb以上就不太好插入了，而且效率也低，做老鼠的话打那
: 么多embryo，还要都生出来做genotyping,都是钱啊... 鱼的我不懂，可能便宜很多吧
G*m
119 楼
因为个儿小，做neural circuit很有优势，能比较容易看到整个功能区的细胞活动，不
像老鼠只见树木不见森林。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
像老鼠只见树木不见森林。
model
transpare
rep
to
很多
serious的
大好
【在 p*****m 的大作中提到】
: 刚刚学了学怎么伺候鱼，交配，收集larvae blabla,同时做点分子实验，同时了解了解
: 大家都在干啥。基本还没入门。不过感觉和之前还是稍微有点不同的 对fish这个model
: （只考虑非发育生物学的应用，这个做的已经没太大意思了）
: small molecule screen还是仍旧给力。如果做能够take advantage of its transpare
: ncy的东西也很给力，比如看看血液循环，心脏搏动之类的的，能用到fluorescent rep
: orter的也不错（但是看下面）
: 同时开始觉得用fish做genetics不是那么爽了。做stable transgene时间太长，而且to
: olkit比起果蝇和小鼠差得太远，不光是大类的工具不行，就是有的工具也需要很多很多
: refinement才能到老鼠果蝇的完善程度（有些在改进），我现在开始觉得如果serious的
: 做fish genetics(forward or reverse)还不如小鼠来的爽。同时，fish吸引我的一大好
A*A
120 楼
目前的观点是，HR和NHEJ是在DNA双链断裂修复中互相竞争的pathways。如果能够有效
的抑制NHEJ,HR的频率有可能会有大幅度的提升。目前，一些labs主要通过knock out或
者knock down NHEJ中几个关键player， 比如，Ku70， Ku80 以及Ligase4，来进行验
证。比较convincing的data有Dana Carroll在果蝇的研究，以及Eric Hendrickson在
mammalian cell line中的研究。Paper 如下:
Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8. Epub 2008 Jun 18.
Ku70, an essential gene, modulates the frequency of rAAV-mediated gene
targeting in human somatic cells.
Fattah FJ, Lichter NF, Fattah KR, Oh S, Hendrickson EA.
Genetics. 2009 Jul;182(3):641-51. Epub 2009 Apr 20.
Genetic analysis of zinc-finger nuclease-induced gene targeting in
Drosophila.
Bozas A, Beumer KJ, Trautman JK, Carroll D.
不过，细胞究竟会使用什么pathway修复双链断裂应该复杂得多。至少细胞周期和发育
阶段也会对pathway的选择有很大影响。我曾经和一个做fish的PI聊过为什么ZFN-
mediated HR在fish里的效果这么不理想。他的看法是fish的embryo发育太快，所以一
旦有双链断裂需要修复，细胞会采取最快捷的NHEJ来修复，而不是需要更多时间和能量
的HR.
【在 p*****m 的大作中提到】
: 嗯 这几天刚有paper出来show这个的。请问一下有什么具体思路来一直NHEJ,然后这个
: 为什么能促进HR呢？我对这些分子生物学东东一窍不通 要是有相关文献就更好了哈 多
: 谢多谢
:
: sequence
: 率。
的抑制NHEJ,HR的频率有可能会有大幅度的提升。目前，一些labs主要通过knock out或
者knock down NHEJ中几个关键player， 比如，Ku70， Ku80 以及Ligase4，来进行验
证。比较convincing的data有Dana Carroll在果蝇的研究，以及Eric Hendrickson在
mammalian cell line中的研究。Paper 如下:
Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8. Epub 2008 Jun 18.
Ku70, an essential gene, modulates the frequency of rAAV-mediated gene
targeting in human somatic cells.
Fattah FJ, Lichter NF, Fattah KR, Oh S, Hendrickson EA.
Genetics. 2009 Jul;182(3):641-51. Epub 2009 Apr 20.
Genetic analysis of zinc-finger nuclease-induced gene targeting in
Drosophila.
Bozas A, Beumer KJ, Trautman JK, Carroll D.
不过，细胞究竟会使用什么pathway修复双链断裂应该复杂得多。至少细胞周期和发育
阶段也会对pathway的选择有很大影响。我曾经和一个做fish的PI聊过为什么ZFN-
mediated HR在fish里的效果这么不理想。他的看法是fish的embryo发育太快，所以一
旦有双链断裂需要修复，细胞会采取最快捷的NHEJ来修复，而不是需要更多时间和能量
的HR.
【在 p*****m 的大作中提到】
: 嗯 这几天刚有paper出来show这个的。请问一下有什么具体思路来一直NHEJ,然后这个
: 为什么能促进HR呢？我对这些分子生物学东东一窍不通 要是有相关文献就更好了哈 多
: 谢多谢
:
: sequence
: 率。
G*m
121 楼
哈，这个主题帖很多给力的内容
【在 A****A 的大作中提到】
: 目前的观点是，HR和NHEJ是在DNA双链断裂修复中互相竞争的pathways。如果能够有效
: 的抑制NHEJ,HR的频率有可能会有大幅度的提升。目前，一些labs主要通过knock out或
: 者knock down NHEJ中几个关键player， 比如，Ku70， Ku80 以及Ligase4，来进行验
: 证。比较convincing的data有Dana Carroll在果蝇的研究，以及Eric Hendrickson在
: mammalian cell line中的研究。Paper 如下:
: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8. Epub 2008 Jun 18.
: Ku70, an essential gene, modulates the frequency of rAAV-mediated gene
: targeting in human somatic cells.
: Fattah FJ, Lichter NF, Fattah KR, Oh S, Hendrickson EA.
: Genetics. 2009 Jul;182(3):641-51. Epub 2009 Apr 20.
【在 A****A 的大作中提到】
: 目前的观点是，HR和NHEJ是在DNA双链断裂修复中互相竞争的pathways。如果能够有效
: 的抑制NHEJ,HR的频率有可能会有大幅度的提升。目前，一些labs主要通过knock out或
: 者knock down NHEJ中几个关键player， 比如，Ku70， Ku80 以及Ligase4，来进行验
: 证。比较convincing的data有Dana Carroll在果蝇的研究，以及Eric Hendrickson在
: mammalian cell line中的研究。Paper 如下:
: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8. Epub 2008 Jun 18.
: Ku70, an essential gene, modulates the frequency of rAAV-mediated gene
: targeting in human somatic cells.
: Fattah FJ, Lichter NF, Fattah KR, Oh S, Hendrickson EA.
: Genetics. 2009 Jul;182(3):641-51. Epub 2009 Apr 20.
f*u
122 楼
double stranded DNA break会增加HR的效率，这个应该是一个普遍现象。
所以对于ZFN来说，增加HR这个根本不需要去做，因为它本身就在那里。
我觉得可能是鱼的重组效率实在太低，就算诱导了DNA break也还是很低。
【在 p*****m 的大作中提到】
: ZFN在鱼里目前就是用来造成NHEJ来形成mutation的 你说这个增加HR效率的事情 目前
: 好像还停留在我脑子里 当然也许已经有人开做了:)
所以对于ZFN来说，增加HR这个根本不需要去做，因为它本身就在那里。
我觉得可能是鱼的重组效率实在太低，就算诱导了DNA break也还是很低。
【在 p*****m 的大作中提到】
: ZFN在鱼里目前就是用来造成NHEJ来形成mutation的 你说这个增加HR效率的事情 目前
: 好像还停留在我脑子里 当然也许已经有人开做了:)
p*m
123 楼
我知道你的意思 我是说没人在fish里真的去好好试用ZFN做HR
【在 f**u 的大作中提到】
: double stranded DNA break会增加HR的效率，这个应该是一个普遍现象。
: 所以对于ZFN来说，增加HR这个根本不需要去做，因为它本身就在那里。
: 我觉得可能是鱼的重组效率实在太低，就算诱导了DNA break也还是很低。
【在 f**u 的大作中提到】
: double stranded DNA break会增加HR的效率，这个应该是一个普遍现象。
: 所以对于ZFN来说，增加HR这个根本不需要去做，因为它本身就在那里。
: 我觉得可能是鱼的重组效率实在太低，就算诱导了DNA break也还是很低。
f*u
124 楼
如果原因真的是像你提到的这个PI说得这样的话，可以试试germline transformation
【在 A****A 的大作中提到】
: 目前的观点是，HR和NHEJ是在DNA双链断裂修复中互相竞争的pathways。如果能够有效
: 的抑制NHEJ,HR的频率有可能会有大幅度的提升。目前，一些labs主要通过knock out或
: 者knock down NHEJ中几个关键player， 比如，Ku70， Ku80 以及Ligase4，来进行验
: 证。比较convincing的data有Dana Carroll在果蝇的研究，以及Eric Hendrickson在
: mammalian cell line中的研究。Paper 如下:
: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8. Epub 2008 Jun 18.
: Ku70, an essential gene, modulates the frequency of rAAV-mediated gene
: targeting in human somatic cells.
: Fattah FJ, Lichter NF, Fattah KR, Oh S, Hendrickson EA.
: Genetics. 2009 Jul;182(3):641-51. Epub 2009 Apr 20.
【在 A****A 的大作中提到】
: 目前的观点是，HR和NHEJ是在DNA双链断裂修复中互相竞争的pathways。如果能够有效
: 的抑制NHEJ,HR的频率有可能会有大幅度的提升。目前，一些labs主要通过knock out或
: 者knock down NHEJ中几个关键player， 比如，Ku70， Ku80 以及Ligase4，来进行验
: 证。比较convincing的data有Dana Carroll在果蝇的研究，以及Eric Hendrickson在
: mammalian cell line中的研究。Paper 如下:
: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8. Epub 2008 Jun 18.
: Ku70, an essential gene, modulates the frequency of rAAV-mediated gene
: targeting in human somatic cells.
: Fattah FJ, Lichter NF, Fattah KR, Oh S, Hendrickson EA.
: Genetics. 2009 Jul;182(3):641-51. Epub 2009 Apr 20.
x*w
125 楼
ＵC Berkeley的Sharon Amacher 2010年fish meeting上就有post说在抑制NHEJ的条
件下促进hr的尝试，当时已经拿到heters 了，搞不好过两天文章就出来了，大家还在
这里扯。。。。呵呵。。。
transformation
【在 f**u 的大作中提到】
: 如果原因真的是像你提到的这个PI说得这样的话，可以试试germline transformation
件下促进hr的尝试，当时已经拿到heters 了，搞不好过两天文章就出来了，大家还在
这里扯。。。。呵呵。。。
transformation
【在 f**u 的大作中提到】
: 如果原因真的是像你提到的这个PI说得这样的话，可以试试germline transformation
i*g
126 楼
NHEJ HR
我2003年就接触了，也是个焦头烂额的领域
发文章是可以的
但要指望它做些事情，提高HR之类的，这个叫产品，或工具。从文章到产品 还有一段路
生物系统，凡是卷入N个蛋白，别指望容易的事情
我2003年就接触了，也是个焦头烂额的领域
发文章是可以的
但要指望它做些事情，提高HR之类的，这个叫产品，或工具。从文章到产品 还有一段路
生物系统，凡是卷入N个蛋白，别指望容易的事情
m*5
127 楼
老鼠可以体外受精阿……JAX lab rescue老line就用这个的，5000刀一个Line
还有用IVF高速expand所需要的line
【在 O******e 的大作中提到】
: 以后我如果有闲工夫就搞搞这个老鼠的体外受精，呵呵
还有用IVF高速expand所需要的line
【在 O******e 的大作中提到】
: 以后我如果有闲工夫就搞搞这个老鼠的体外受精，呵呵
m*5
128 楼
老鼠Knock in打的是mES cell,挑 colony,体力活，但是不难
【在 G*****m 的大作中提到】
: 插个loxP,attP之类的就很好用了。。。
【在 G*****m 的大作中提到】
: 插个loxP,attP之类的就很好用了。。。
X*n
129 楼
我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
p*m
130 楼
你说的我赶紧去翻了一下abs book和笔记，他们好像没提成功了啊 去年的会上有好几
家号称在用ZFN直接做KI，没有一个说自己看到成功了。
【在 x**w 的大作中提到】
: ＵC Berkeley的Sharon Amacher 2010年fish meeting上就有post说在抑制NHEJ的条
: 件下促进hr的尝试，当时已经拿到heters 了，搞不好过两天文章就出来了，大家还在
: 这里扯。。。。呵呵。。。
:
: transformation
家号称在用ZFN直接做KI，没有一个说自己看到成功了。
【在 x**w 的大作中提到】
: ＵC Berkeley的Sharon Amacher 2010年fish meeting上就有post说在抑制NHEJ的条
: 件下促进hr的尝试，当时已经拿到heters 了，搞不好过两天文章就出来了，大家还在
: 这里扯。。。。呵呵。。。
:
: transformation
s*r
131 楼
再仔细讲讲嘛。
【在 X******n 的大作中提到】
: 我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
【在 X******n 的大作中提到】
: 我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
X*n
132 楼
我问了一下，他说不是在fish里做的。我搞错了。
【在 s********r 的大作中提到】
: 再仔细讲讲嘛。
【在 s********r 的大作中提到】
: 再仔细讲讲嘛。
x*w
133 楼
我又去翻了下我的笔记。。呵呵。。
更正一下，是拿到了chimera fish
【在 p*****m 的大作中提到】
: 你说的我赶紧去翻了一下abs book和笔记，他们好像没提成功了啊 去年的会上有好几
: 家号称在用ZFN直接做KI，没有一个说自己看到成功了。
更正一下，是拿到了chimera fish
【在 p*****m 的大作中提到】
: 你说的我赶紧去翻了一下abs book和笔记，他们好像没提成功了啊 去年的会上有好几
: 家号称在用ZFN直接做KI，没有一个说自己看到成功了。
f*u
134 楼
这个嘛，关键看能不能把germline给KI了。
【在 x**w 的大作中提到】
: 我又去翻了下我的笔记。。呵呵。。
: 更正一下，是拿到了chimera fish
【在 x**w 的大作中提到】
: 我又去翻了下我的笔记。。呵呵。。
: 更正一下，是拿到了chimera fish
c*m
135 楼
knock-in 和 knock-out 有啥区别阿？
【在 M*****n 的大作中提到】
: 俺对ZFN这个field知道不多,
: 好像作knock-in，还是要靠hom rec的， 虽然ZFN可以有所帮助。
:
: 要的
【在 M*****n 的大作中提到】
: 俺对ZFN这个field知道不多,
: 好像作knock-in，还是要靠hom rec的， 虽然ZFN可以有所帮助。
:
: 要的
M*n
136 楼
老鼠还是鱼？
rat去年就发到nature biotech上了。
【在 X******n 的大作中提到】
: 我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
rat去年就发到nature biotech上了。
【在 X******n 的大作中提到】
: 我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
M*n
137 楼
下一步就是要拿到germline
光是chimera 还发不了大paper.
其实HR肯定可以发生，就是效率太低。
俺有一个想法就是
如果直接manipulate embryos效率太低的话，可以先转一个artificial chromosome作
一个stableline.然后跟抑制NHJR的mutant line cross，然后后代筛选homologous
recombinants.
鱼能产很多卵，筛选几万个应该不难。
光是chimera 还发不了大paper.
其实HR肯定可以发生，就是效率太低。
俺有一个想法就是
如果直接manipulate embryos效率太低的话，可以先转一个artificial chromosome作
一个stableline.然后跟抑制NHJR的mutant line cross，然后后代筛选homologous
recombinants.
鱼能产很多卵，筛选几万个应该不难。
p*r
138 楼
我最近在研究全老鼠的imaging...
技术上最近还是颇有些新方法的， 我觉得以后看个胚胎应该是可以的。
【在 p*****m 的大作中提到】
: 你这个冷水。。。鱼透明这个好处是替代不了的 而且人家体外受精 你们老鼠行么。。
: 呵呵
技术上最近还是颇有些新方法的， 我觉得以后看个胚胎应该是可以的。
【在 p*****m 的大作中提到】
: 你这个冷水。。。鱼透明这个好处是替代不了的 而且人家体外受精 你们老鼠行么。。
: 呵呵
p*r
139 楼
TALE 其实还暂时没那么好。
【在 A****A 的大作中提到】
: 具我所知，几个fish labs正在联合用ZFN搞大规模的knock out resources。ZFN以及相
: 关技术最近会有比较大的进展。大家如果感兴趣，可以关注一下最近的TALEN技术。在
: 最近1-2年内，估计任何试验室都可以自己合成reagent去target任何基因。cost不会比
: 买限制性内切酶高太多。
:
: model
: transpare
: rep
: to
: 很多
【在 A****A 的大作中提到】
: 具我所知，几个fish labs正在联合用ZFN搞大规模的knock out resources。ZFN以及相
: 关技术最近会有比较大的进展。大家如果感兴趣，可以关注一下最近的TALEN技术。在
: 最近1-2年内，估计任何试验室都可以自己合成reagent去target任何基因。cost不会比
: 买限制性内切酶高太多。
:
: model
: transpare
: rep
: to
: 很多
p*r
140 楼
但是 fish靠谱的behavior不多， 现在也就主要有人搞点motor behavior.
【在 G*****m 的大作中提到】
: 因为个儿小，做neural circuit很有优势，能比较容易看到整个功能区的细胞活动，不
: 像老鼠只见树木不见森林。
:
: model
: transpare
: rep
: to
: 很多
: serious的
: 大好
【在 G*****m 的大作中提到】
: 因为个儿小，做neural circuit很有优势，能比较容易看到整个功能区的细胞活动，不
: 像老鼠只见树木不见森林。
:
: model
: transpare
: rep
: to
: 很多
: serious的
: 大好
s*y
141 楼
能看到什么样的分辨率和深度？
可以看到荧光么？
【在 p*******r 的大作中提到】
: 我最近在研究全老鼠的imaging...
: 技术上最近还是颇有些新方法的， 我觉得以后看个胚胎应该是可以的。
可以看到荧光么？
【在 p*******r 的大作中提到】
: 我最近在研究全老鼠的imaging...
: 技术上最近还是颇有些新方法的， 我觉得以后看个胚胎应该是可以的。
D*a
142 楼
胚胎不是在老鼠肚子里么？
【在 p*******r 的大作中提到】
: 我最近在研究全老鼠的imaging...
: 技术上最近还是颇有些新方法的， 我觉得以后看个胚胎应该是可以的。
【在 p*******r 的大作中提到】
: 我最近在研究全老鼠的imaging...
: 技术上最近还是颇有些新方法的， 我觉得以后看个胚胎应该是可以的。
D*a
143 楼
这个理论上是confidential吧，还是删了吧。
【在 X******n 的大作中提到】
: 我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
【在 X******n 的大作中提到】
: 我好像看到老板在审一篇用ＺＦＮ做Ｋｎｏｃｋｉｎ的文章，发到Ｎａｔｕｒｅ上的。
p*m
144 楼
最近很有几篇TALEN的应用 看起来效率至少不比ZFN差啊 programability更是远胜
【在 p*******r 的大作中提到】
: TALE 其实还暂时没那么好。
【在 p*******r 的大作中提到】
: TALE 其实还暂时没那么好。
p*m
145 楼
这个我觉得是搞的人不多 而不是靠谱的行为不多
【在 p*******r 的大作中提到】
: 但是 fish靠谱的behavior不多， 现在也就主要有人搞点motor behavior.
【在 p*******r 的大作中提到】
: 但是 fish靠谱的behavior不多， 现在也就主要有人搞点motor behavior.
w*s
146 楼
How about stem cell research in fish? Isn't that part of the developmental
biology?
【在 M*****n 的大作中提到】
: 俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
: 你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
: developmental biology is pretty much over. don't waster your time on it.
: pushing/shaking flies is good for beginners to learn genetics.
: my 2 cents.
:
: model
: transpare
: rep
: to
biology?
【在 M*****n 的大作中提到】
: 俺昨天不是说了，next big thing 就是knockout fish. 你应该朝这个方向努力。如果
: 你把这个做出来，虽然得不了炸药将，但是名气也不会小了。
: developmental biology is pretty much over. don't waster your time on it.
: pushing/shaking flies is good for beginners to learn genetics.
: my 2 cents.
:
: model
: transpare
: rep
: to
p*r
147 楼
哦， 似乎off target要多一点。
【在 p*****m 的大作中提到】
: 最近很有几篇TALEN的应用 看起来效率至少不比ZFN差啊 programability更是远胜
【在 p*****m 的大作中提到】
: 最近很有几篇TALEN的应用 看起来效率至少不比ZFN差啊 programability更是远胜
A*A
148 楼
实际上，目前大部分证据表明TALEN的off target效应要小于ZFN. TALEN现在的主要缺
点在于size太大，一个TALEN coding sequecne 至少要4kb, 而ZFN则只要1kb左右。而
且从目前以发表的和我们自己的数据来看，TALEN的剪切活性可能要低于ZFN。现在一些
实验室都在进一步优化TALEN的size以及活性，希望以后能有大幅度的提高。TALEN相对
于ZFN目前最大的优势是位点的选择范围更大。基本上每5-10bp就可以找到一个好的
TALEN位点。ZFN目前是200bp-500bp. 另外TALEN的出现打破了ZFN在genome
engineering领域的垄断地位。将大大降低reagent的价格。现在法国的Cellectis就提
供$5000TALEN的订制服务。Sigma的ZFN价格也会跟着降下来。
【在 p*******r 的大作中提到】
: 哦， 似乎off target要多一点。
点在于size太大，一个TALEN coding sequecne 至少要4kb, 而ZFN则只要1kb左右。而
且从目前以发表的和我们自己的数据来看，TALEN的剪切活性可能要低于ZFN。现在一些
实验室都在进一步优化TALEN的size以及活性，希望以后能有大幅度的提高。TALEN相对
于ZFN目前最大的优势是位点的选择范围更大。基本上每5-10bp就可以找到一个好的
TALEN位点。ZFN目前是200bp-500bp. 另外TALEN的出现打破了ZFN在genome
engineering领域的垄断地位。将大大降低reagent的价格。现在法国的Cellectis就提
供$5000TALEN的订制服务。Sigma的ZFN价格也会跟着降下来。
【在 p*******r 的大作中提到】
: 哦， 似乎off target要多一点。
u*d
149 楼
talen一定要那么多的手指吗？手指多了off target会不会反而多啊？
【在 A****A 的大作中提到】
: 实际上，目前大部分证据表明TALEN的off target效应要小于ZFN. TALEN现在的主要缺
: 点在于size太大，一个TALEN coding sequecne 至少要4kb, 而ZFN则只要1kb左右。而
: 且从目前以发表的和我们自己的数据来看，TALEN的剪切活性可能要低于ZFN。现在一些
: 实验室都在进一步优化TALEN的size以及活性，希望以后能有大幅度的提高。TALEN相对
: 于ZFN目前最大的优势是位点的选择范围更大。基本上每5-10bp就可以找到一个好的
: TALEN位点。ZFN目前是200bp-500bp. 另外TALEN的出现打破了ZFN在genome
: engineering领域的垄断地位。将大大降低reagent的价格。现在法国的Cellectis就提
: 供$5000TALEN的订制服务。Sigma的ZFN价格也会跟着降下来。
【在 A****A 的大作中提到】
: 实际上，目前大部分证据表明TALEN的off target效应要小于ZFN. TALEN现在的主要缺
: 点在于size太大，一个TALEN coding sequecne 至少要4kb, 而ZFN则只要1kb左右。而
: 且从目前以发表的和我们自己的数据来看，TALEN的剪切活性可能要低于ZFN。现在一些
: 实验室都在进一步优化TALEN的size以及活性，希望以后能有大幅度的提高。TALEN相对
: 于ZFN目前最大的优势是位点的选择范围更大。基本上每5-10bp就可以找到一个好的
: TALEN位点。ZFN目前是200bp-500bp. 另外TALEN的出现打破了ZFN在genome
: engineering领域的垄断地位。将大大降低reagent的价格。现在法国的Cellectis就提
: 供$5000TALEN的订制服务。Sigma的ZFN价格也会跟着降下来。
k*o
150 楼
目前各大pharma用in vitro和cell-based的平台都筛不出好的candidate molecule。这
样的形势下，用鱼做screen到底有多大意义？
样的形势下，用鱼做screen到底有多大意义？
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