待遇跟前面承诺的差一大截， 咋办？(ZZ)# Chemistry - 化学
n*n
1 楼
爸妈打算十月过来探亲，符合中信代签，但是看了一些帖子，有些问题想请教一下
有人说要填DS160，还需要填156和157么
有人说要交邀请信，有人没有交也过了，这个是必需的么
有没有比较全面的帖子说那些是必须的材料啊，谢谢了
有人说要填DS160，还需要填156和157么
有人说要交邀请信，有人没有交也过了，这个是必需的么
有没有比较全面的帖子说那些是必须的材料啊，谢谢了
s*o
2 楼
ds160里提到是否受过专门训练，生物，化学之类，这个问题应该说
Yes 还是 NO？
Yes 还是 NO？
t*r
3 楼
从看真正男子汉的时候看到王宝强的儿子，当时就觉得这他妈完全不一样啊！当时我还感叹王宝强他媳妇马蓉的基因还算强大，生了这么个小帅哥，但是王宝强公然公开自己被戴绿帽子了！！
快去看看王宝强的儿子和宋喆，他妈的神情五官……王宝强的媳妇儿马蓉和宋喆出轨绝对不是几个月的事！傻宝宝，你媳妇儿估计在一开始和你的婚姻就是一场阴谋啊！估计那时候就已经和宋喆勾搭上了，可怜的宝宝，结婚这么多年你一直在和别人共享妻子啊！不对共享性伴侣。
我一直觉得王宝强离婚这个事儿，不仅仅是马蓉红杏出墙这么简单，别问我为啥有这种感觉，只能说女人的第六感简直不能他妈的再准了。单单从王宝强小儿子和怂喆如出一辙的事来看，很可能是马蓉和宋喆发生关系怀了王宝强的“儿子”，王宝强顶包，俩人集体串通搜刮王宝强的钱财，一步步得逞，现在他妈的东窗事发了。
离婚，必须离婚，不离婚就不是真男人了。而且我相信，出轨有证据的情况下，孩子绝对不会给荡妇的！不过，女儿收了，儿子就算了吧（强烈建议去做一下DNA，没别的意思），别再给宋喆养孩子了！
快去看看王宝强的儿子和宋喆，他妈的神情五官……王宝强的媳妇儿马蓉和宋喆出轨绝对不是几个月的事！傻宝宝，你媳妇儿估计在一开始和你的婚姻就是一场阴谋啊！估计那时候就已经和宋喆勾搭上了，可怜的宝宝，结婚这么多年你一直在和别人共享妻子啊！不对共享性伴侣。
我一直觉得王宝强离婚这个事儿，不仅仅是马蓉红杏出墙这么简单，别问我为啥有这种感觉，只能说女人的第六感简直不能他妈的再准了。单单从王宝强小儿子和怂喆如出一辙的事来看，很可能是马蓉和宋喆发生关系怀了王宝强的“儿子”，王宝强顶包，俩人集体串通搜刮王宝强的钱财，一步步得逞，现在他妈的东窗事发了。
离婚，必须离婚，不离婚就不是真男人了。而且我相信，出轨有证据的情况下，孩子绝对不会给荡妇的！不过，女儿收了，儿子就算了吧（强烈建议去做一下DNA，没别的意思），别再给宋喆养孩子了！
w*e
4 楼
I suspected my dsDNA (100bp) pool (random sequence) dissociate and
form a messy mesh structure. When I run sample on a gel, many bands
show up above the expected molecular weight.
Anybody has exp with this problem? Thanks!
form a messy mesh structure. When I run sample on a gel, many bands
show up above the expected molecular weight.
Anybody has exp with this problem? Thanks!
s*u
5 楼
发信人: aiquanquan (gougou), 信区: Returnee
标 题: 问一个国内高校offer letter里头的说法
发信站: BBS 未名空间站 (Sun Jan 22 13:06:04 2012, 美东)
待遇第一条写着：
工资待遇：初聘教授岗位工资约年薪13万（含个人公积金、保险及税，另440元/月的午
餐补贴发到教工卡中）。
不知道该怎么断句，比如说“初聘教授岗位，工资约年薪13万”，还是“初聘教授，岗
位工资约年薪13万”？我搜出来国内工资单有“岗位工资”这个名目，不知道实际收入
除了这个，有没有其他名目，比如“岗位津贴”等，那就有比较大的上升空间。如果这
13万就是全部，那这待遇跟刚开始说的出入很大，又不知道应该找哪个负责人问清楚，
一直在接洽的那个头以后是直接领导，口头上谈过两次待遇都给了很好的条件，现在逼
人家罗列出来细则又好像比较过分，但我也不想被这样坑回国。有建议么？
谢谢答复。真搞不懂那些个领导，一开始就忽悠人，咱又不是什么人才，最后也占了人
家的名额申请了青千，现在又不能兑现学校这块的待遇，让人进退两难，这又是何苦呢
？！至于中组部那个津贴，写着一次性50万房补，又不同于大家说的每年10万给五年，
真是一头雾水。最后还来个“数字仅供参考”。
中组部的启动经费是100-300万，学校给50万。
现在人家白纸黑子写的待遇跟前面承诺的差一大截，再不如实都兑现了，那真是黑店了。
手头还有interview，不过不一定能成。在美国两博后好死赖活着，跟现在回国一个教
授相比，你选哪个
标 题: 问一个国内高校offer letter里头的说法
发信站: BBS 未名空间站 (Sun Jan 22 13:06:04 2012, 美东)
待遇第一条写着：
工资待遇：初聘教授岗位工资约年薪13万（含个人公积金、保险及税，另440元/月的午
餐补贴发到教工卡中）。
不知道该怎么断句，比如说“初聘教授岗位，工资约年薪13万”，还是“初聘教授，岗
位工资约年薪13万”？我搜出来国内工资单有“岗位工资”这个名目，不知道实际收入
除了这个，有没有其他名目，比如“岗位津贴”等，那就有比较大的上升空间。如果这
13万就是全部，那这待遇跟刚开始说的出入很大，又不知道应该找哪个负责人问清楚，
一直在接洽的那个头以后是直接领导，口头上谈过两次待遇都给了很好的条件，现在逼
人家罗列出来细则又好像比较过分，但我也不想被这样坑回国。有建议么？
谢谢答复。真搞不懂那些个领导，一开始就忽悠人，咱又不是什么人才，最后也占了人
家的名额申请了青千，现在又不能兑现学校这块的待遇，让人进退两难，这又是何苦呢
？！至于中组部那个津贴，写着一次性50万房补，又不同于大家说的每年10万给五年，
真是一头雾水。最后还来个“数字仅供参考”。
中组部的启动经费是100-300万，学校给50万。
现在人家白纸黑子写的待遇跟前面承诺的差一大截，再不如实都兑现了，那真是黑店了。
手头还有interview，不过不一定能成。在美国两博后好死赖活着，跟现在回国一个教
授相比，你选哪个
t*s
6 楼
看置顶的常见问题。
里面有连接。
里面有连接。
s*e
7 楼
这个不可能. 100 bp DNA已经非常稳定了. 100 bp DNA成环后也不会melt, 何况你的是
线性DNA.
【在 w******e 的大作中提到】
: I suspected my dsDNA (100bp) pool (random sequence) dissociate and
: form a messy mesh structure. When I run sample on a gel, many bands
: show up above the expected molecular weight.
: Anybody has exp with this problem? Thanks!
线性DNA.
【在 w******e 的大作中提到】
: I suspected my dsDNA (100bp) pool (random sequence) dissociate and
: form a messy mesh structure. When I run sample on a gel, many bands
: show up above the expected molecular weight.
: Anybody has exp with this problem? Thanks!
x*9
8 楼
换阿，
c*u
9 楼
邀请信，不是很需要
s*e
10 楼
oh. 你这个是100 bp DNA 环吗?
【在 w******e 的大作中提到】
: I suspected my dsDNA (100bp) pool (random sequence) dissociate and
: form a messy mesh structure. When I run sample on a gel, many bands
: show up above the expected molecular weight.
: Anybody has exp with this problem? Thanks!
【在 w******e 的大作中提到】
: I suspected my dsDNA (100bp) pool (random sequence) dissociate and
: form a messy mesh structure. When I run sample on a gel, many bands
: show up above the expected molecular weight.
: Anybody has exp with this problem? Thanks!
l*y
11 楼
有效护照，DS-160确认表，照片，英文邀请信，签证申请费，代办费200元，回传EMS费
50元，三个星期内收到签证。
50元，三个星期内收到签证。
h*6
12 楼
If it is high AT DNA, DNA may melt during gel running. If DNA melts, I
doubt it will re-form dsDNA under the same condition. ssDNA migrate slower.
doubt it will re-form dsDNA under the same condition. ssDNA migrate slower.
s*e
13 楼
如果是环, 100 bp的DNA在这么高的曲率下, 可能会有局部melting (one or a few bp)
来环解弯曲弹性能.在跑胶的温度下, 这样的local melting sites可能会有数个. 不同
的个数会导致DNA不同的构型, gel shift速度会不一样.
【在 s***e 的大作中提到】
: oh. 你这个是100 bp DNA 环吗?
来环解弯曲弹性能.在跑胶的温度下, 这样的local melting sites可能会有数个. 不同
的个数会导致DNA不同的构型, gel shift速度会不一样.
【在 s***e 的大作中提到】
: oh. 你这个是100 bp DNA 环吗?
w*e
14 楼
thx for the info. no it's not circular DNA. it's random pool, so if the
ds structure ever dissociates, there is virtually no chance for
re-annealing thou. I just don't know whether it's possible for the ds
structure to dissociate under normal experimental condition (like gel
running, or salt solution, etc)
bp)
【在 s***e 的大作中提到】
: 如果是环, 100 bp的DNA在这么高的曲率下, 可能会有局部melting (one or a few bp)
: 来环解弯曲弹性能.在跑胶的温度下, 这样的local melting sites可能会有数个. 不同
: 的个数会导致DNA不同的构型, gel shift速度会不一样.
ds structure ever dissociates, there is virtually no chance for
re-annealing thou. I just don't know whether it's possible for the ds
structure to dissociate under normal experimental condition (like gel
running, or salt solution, etc)
bp)
【在 s***e 的大作中提到】
: 如果是环, 100 bp的DNA在这么高的曲率下, 可能会有局部melting (one or a few bp)
: 来环解弯曲弹性能.在跑胶的温度下, 这样的local melting sites可能会有数个. 不同
: 的个数会导致DNA不同的构型, gel shift速度会不一样.
s*e
15 楼
不太可能. 单分子unzip DNA hairpin的测量里, DNA一般比100 bp还短. normal
experimental condition下需要> 10pN才能把双链分开.
【在 w******e 的大作中提到】
: thx for the info. no it's not circular DNA. it's random pool, so if the
: ds structure ever dissociates, there is virtually no chance for
: re-annealing thou. I just don't know whether it's possible for the ds
: structure to dissociate under normal experimental condition (like gel
: running, or salt solution, etc)
:
: bp)
experimental condition下需要> 10pN才能把双链分开.
【在 w******e 的大作中提到】
: thx for the info. no it's not circular DNA. it's random pool, so if the
: ds structure ever dissociates, there is virtually no chance for
: re-annealing thou. I just don't know whether it's possible for the ds
: structure to dissociate under normal experimental condition (like gel
: running, or salt solution, etc)
:
: bp)
a*e
16 楼
make fresh buffer and run the gel again.
make sure the loading buffer is correct.
generally, ssDNA runs faster than dsDNA with the same length. So should run
below, not above, your expected band.
make sure the loading buffer is correct.
generally, ssDNA runs faster than dsDNA with the same length. So should run
below, not above, your expected band.
w*e
17 楼
thx again. my situation is: I gel purified some 100bp pool a while ago.
the pool looked great on the gel for extraction (sharp 100bp band).
I didn't bother to run gel again after the extraction (stupid of me)
and just spec them and threw them in the freezer. I noticed a high
salt peak at 230 but felt I don't have to care for now.
today (~2 months later) I took it out, mix w/ normal loading buffer,
and ran it on a native TBE-PAGE gel as mass standard(I always use this
type of gel for short dsDNA
【在 s***e 的大作中提到】
: 不太可能. 单分子unzip DNA hairpin的测量里, DNA一般比100 bp还短. normal
: experimental condition下需要> 10pN才能把双链分开.
the pool looked great on the gel for extraction (sharp 100bp band).
I didn't bother to run gel again after the extraction (stupid of me)
and just spec them and threw them in the freezer. I noticed a high
salt peak at 230 but felt I don't have to care for now.
today (~2 months later) I took it out, mix w/ normal loading buffer,
and ran it on a native TBE-PAGE gel as mass standard(I always use this
type of gel for short dsDNA
【在 s***e 的大作中提到】
: 不太可能. 单分子unzip DNA hairpin的测量里, DNA一般比100 bp还短. normal
: experimental condition下需要> 10pN才能把双链分开.
s*e
18 楼
我不知道DNA在power form有多稳定. 短DNA出现smear band我见过一次. 不过那是因为
这个DNA里有许多tandem sequence. 所以错位一个周期, 能量不是很大.
【在 w******e 的大作中提到】
: thx again. my situation is: I gel purified some 100bp pool a while ago.
: the pool looked great on the gel for extraction (sharp 100bp band).
: I didn't bother to run gel again after the extraction (stupid of me)
: and just spec them and threw them in the freezer. I noticed a high
: salt peak at 230 but felt I don't have to care for now.
: today (~2 months later) I took it out, mix w/ normal loading buffer,
: and ran it on a native TBE-PAGE gel as mass standard(I always use this
: type of gel for short dsDNA
这个DNA里有许多tandem sequence. 所以错位一个周期, 能量不是很大.
【在 w******e 的大作中提到】
: thx again. my situation is: I gel purified some 100bp pool a while ago.
: the pool looked great on the gel for extraction (sharp 100bp band).
: I didn't bother to run gel again after the extraction (stupid of me)
: and just spec them and threw them in the freezer. I noticed a high
: salt peak at 230 but felt I don't have to care for now.
: today (~2 months later) I took it out, mix w/ normal loading buffer,
: and ran it on a native TBE-PAGE gel as mass standard(I always use this
: type of gel for short dsDNA
w*e
19 楼
thx anyway. I'll report to the board if I found anything interesting:)
【在 s***e 的大作中提到】
: 我不知道DNA在power form有多稳定. 短DNA出现smear band我见过一次. 不过那是因为
: 这个DNA里有许多tandem sequence. 所以错位一个周期, 能量不是很大.
【在 s***e 的大作中提到】
: 我不知道DNA在power form有多稳定. 短DNA出现smear band我见过一次. 不过那是因为
: 这个DNA里有许多tandem sequence. 所以错位一个周期, 能量不是很大.
a*o
20 楼
I did not read all posts. I guess LZ is doing SELEX. What he/she refered is
the bulge structure formed by many different molecules share two constant
regions at both ends, but with different sequences in the middle.
The simplest way to solve this problem is to dilute your sample to 4 fold
after PCR, add do ONE more cycle of PCR.
the bulge structure formed by many different molecules share two constant
regions at both ends, but with different sequences in the middle.
The simplest way to solve this problem is to dilute your sample to 4 fold
after PCR, add do ONE more cycle of PCR.
w*e
21 楼
thx for the input. PCR is a great idea. so did you see this kind of
dissociation happening in your exp? what are the factors that affect
the process? storage buffer? time? it's surprising to me as all the
newly prepared dsDNA pool have no such problem on gel.
is
【在 a****o 的大作中提到】
: I did not read all posts. I guess LZ is doing SELEX. What he/she refered is
: the bulge structure formed by many different molecules share two constant
: regions at both ends, but with different sequences in the middle.
: The simplest way to solve this problem is to dilute your sample to 4 fold
: after PCR, add do ONE more cycle of PCR.
dissociation happening in your exp? what are the factors that affect
the process? storage buffer? time? it's surprising to me as all the
newly prepared dsDNA pool have no such problem on gel.
is
【在 a****o 的大作中提到】
: I did not read all posts. I guess LZ is doing SELEX. What he/she refered is
: the bulge structure formed by many different molecules share two constant
: regions at both ends, but with different sequences in the middle.
: The simplest way to solve this problem is to dilute your sample to 4 fold
: after PCR, add do ONE more cycle of PCR.
a*o
22 楼
Yes, if you overdid PCR, you will see this phenomenon. Doing PCR for a
fewer cycles may cure the problem. I don't think it is dissociation. When
primer/dNTP/active Taq goes down, annealed DNA-Primer can not be
formed/extended. DNA molecules with different sequence in middle form
duplex. Since they have identical sequences at both ends due to constant
regions, but different sequences in the middle, they can not form perfect
duplex, rather they form partial duplex at both ends, while loop-out
sing
【在 w******e 的大作中提到】
: thx for the input. PCR is a great idea. so did you see this kind of
: dissociation happening in your exp? what are the factors that affect
: the process? storage buffer? time? it's surprising to me as all the
: newly prepared dsDNA pool have no such problem on gel.
:
: is
fewer cycles may cure the problem. I don't think it is dissociation. When
primer/dNTP/active Taq goes down, annealed DNA-Primer can not be
formed/extended. DNA molecules with different sequence in middle form
duplex. Since they have identical sequences at both ends due to constant
regions, but different sequences in the middle, they can not form perfect
duplex, rather they form partial duplex at both ends, while loop-out
sing
【在 w******e 的大作中提到】
: thx for the input. PCR is a great idea. so did you see this kind of
: dissociation happening in your exp? what are the factors that affect
: the process? storage buffer? time? it's surprising to me as all the
: newly prepared dsDNA pool have no such problem on gel.
:
: is
w*e
23 楼
thx a lot for the info. In fact, the product was after PCR optimization
and ran fine on the gel before band cutting and gel extraction, so I
don't think PCR bias is the reason. However, I do agree doing one more
round of PCR is the way to go. If that doesn't get s**t together, there
must be something really wrong with my hands...
【在 a****o 的大作中提到】
: Yes, if you overdid PCR, you will see this phenomenon. Doing PCR for a
: fewer cycles may cure the problem. I don't think it is dissociation. When
: primer/dNTP/active Taq goes down, annealed DNA-Primer can not be
: formed/extended. DNA molecules with different sequence in middle form
: duplex. Since they have identical sequences at both ends due to constant
: regions, but different sequences in the middle, they can not form perfect
: duplex, rather they form partial duplex at both ends, while loop-out
: sing
and ran fine on the gel before band cutting and gel extraction, so I
don't think PCR bias is the reason. However, I do agree doing one more
round of PCR is the way to go. If that doesn't get s**t together, there
must be something really wrong with my hands...
【在 a****o 的大作中提到】
: Yes, if you overdid PCR, you will see this phenomenon. Doing PCR for a
: fewer cycles may cure the problem. I don't think it is dissociation. When
: primer/dNTP/active Taq goes down, annealed DNA-Primer can not be
: formed/extended. DNA molecules with different sequence in middle form
: duplex. Since they have identical sequences at both ends due to constant
: regions, but different sequences in the middle, they can not form perfect
: duplex, rather they form partial duplex at both ends, while loop-out
: sing
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