x*e
2 楼
据版上各位分子克隆大神说Gibson百发百中,正好我要克隆几个比较大的enhancer到9k
的质粒里,就试了一下。positive control连接转化没问题,但是我自己设计引物p出来
的片段跟载体死活连不上。是不是我的引物设计有问题?
的质粒里,就试了一下。positive control连接转化没问题,但是我自己设计引物p出来
的片段跟载体死活连不上。是不是我的引物设计有问题?
d*y
3 楼
Heavy whipping cream打发即可。
B*n
5 楼
1 类似戚风的方子会比海绵蛋糕的不容易裂
2 卷的时候要先轻轻切一刀
3 烤完拿出来要晾一下
这个香港大妈的做法挺好的,可以看下
http://www.youtube.com/watch?v=1uZycFBUHbk
【在 h*******1 的大作中提到】
: 我为什么把长长的蛋糕一卷就裂开?有tip吗?谢谢
2 卷的时候要先轻轻切一刀
3 烤完拿出来要晾一下
这个香港大妈的做法挺好的,可以看下
http://www.youtube.com/watch?v=1uZycFBUHbk
【在 h*******1 的大作中提到】
: 我为什么把长长的蛋糕一卷就裂开?有tip吗?谢谢
h*1
7 楼
谢谢妹子
【在 B********n 的大作中提到】
: 1 类似戚风的方子会比海绵蛋糕的不容易裂
: 2 卷的时候要先轻轻切一刀
: 3 烤完拿出来要晾一下
: 这个香港大妈的做法挺好的,可以看下
: http://www.youtube.com/watch?v=1uZycFBUHbk
【在 B********n 的大作中提到】
: 1 类似戚风的方子会比海绵蛋糕的不容易裂
: 2 卷的时候要先轻轻切一刀
: 3 烤完拿出来要晾一下
: 这个香港大妈的做法挺好的,可以看下
: http://www.youtube.com/watch?v=1uZycFBUHbk
M*g
11 楼
I've used Gibson Assembly more than twenty times and had no problem at all.
Tell me more details. I might be able to help.
how did you open up your vector? PCR or restriction enzymes?
It is better if you purify the pcr product.
don't use competent cells other than NEB. for some reason, it didn't work
well if use invitrogen cells etc.
Dh5alpha comp cell from NEB is better than NEB10 beta regarding
transformation efficiency.
Tell me more details. I might be able to help.
how did you open up your vector? PCR or restriction enzymes?
It is better if you purify the pcr product.
don't use competent cells other than NEB. for some reason, it didn't work
well if use invitrogen cells etc.
Dh5alpha comp cell from NEB is better than NEB10 beta regarding
transformation efficiency.
w*e
12 楼
based on NEB website, it seems neb 10 beta is better than 5 alpha
.
【在 M******g 的大作中提到】
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
: Dh5alpha comp cell from NEB is better than NEB10 beta regarding
: transformation efficiency.
.
【在 M******g 的大作中提到】
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
: Dh5alpha comp cell from NEB is better than NEB10 beta regarding
: transformation efficiency.
w*e
13 楼
我原来做头几次也没问题
后来做复杂的质粒就出问题了,效率很低
请你来讲讲你的经验呀
I've used Gibson Assembly more than twenty times and had no problem at all.
Tell me more details. I might be able to help.
how did you open up your vector? PCR or restriction enzymes?
It is better if you purify the pcr product.
don't use competent cells other than NEB. for some reason, it didn't work
well if use invitrogen cells etc.
Dh5alpha comp cell from NEB is better than NEB10 beta regarding
transformation efficiency.
【在 M******g 的大作中提到】
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
: Dh5alpha comp cell from NEB is better than NEB10 beta regarding
: transformation efficiency.
后来做复杂的质粒就出问题了,效率很低
请你来讲讲你的经验呀
I've used Gibson Assembly more than twenty times and had no problem at all.
Tell me more details. I might be able to help.
how did you open up your vector? PCR or restriction enzymes?
It is better if you purify the pcr product.
don't use competent cells other than NEB. for some reason, it didn't work
well if use invitrogen cells etc.
Dh5alpha comp cell from NEB is better than NEB10 beta regarding
transformation efficiency.
【在 M******g 的大作中提到】
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
: Dh5alpha comp cell from NEB is better than NEB10 beta regarding
: transformation efficiency.
w*e
14 楼
接着这个问个问题
现在life technology也有geneart seamless cloning kit,clontech公司也有
infusion cloning kit,我个人感觉本质上这些kits和NEB gbison kits是一样的
不知道版上面有没有人比较过这几个,那个更稳定
.
【在 w*e 的大作中提到】
: 我原来做头几次也没问题
: 后来做复杂的质粒就出问题了,效率很低
: 请你来讲讲你的经验呀
:
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
现在life technology也有geneart seamless cloning kit,clontech公司也有
infusion cloning kit,我个人感觉本质上这些kits和NEB gbison kits是一样的
不知道版上面有没有人比较过这几个,那个更稳定
.
【在 w*e 的大作中提到】
: 我原来做头几次也没问题
: 后来做复杂的质粒就出问题了,效率很低
: 请你来讲讲你的经验呀
:
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
M*g
19 楼
Tell me more about your 复杂的质粒. I had no problem so far.
.
【在 w*e 的大作中提到】
: 我原来做头几次也没问题
: 后来做复杂的质粒就出问题了,效率很低
: 请你来讲讲你的经验呀
:
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
.
【在 w*e 的大作中提到】
: 我原来做头几次也没问题
: 后来做复杂的质粒就出问题了,效率很低
: 请你来讲讲你的经验呀
:
: I've used Gibson Assembly more than twenty times and had no problem at all.
: Tell me more details. I might be able to help.
: how did you open up your vector? PCR or restriction enzymes?
: It is better if you purify the pcr product.
: don't use competent cells other than NEB. for some reason, it didn't work
: well if use invitrogen cells etc.
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