C*********m
发帖数: 213 | 1 做质粒转化，克隆，质粒比较大，而且带有Tn7, Cre/LoxP sequences. Protocol上建
议用top10,不知道用DH5alpha会不会有影响？多谢。 |
s********r
发帖数: 312 | 2 dont know your case. But we use that constantly for most of the molecular
biology work, including large targeting vectors. They work perfectly. |
C*********m
发帖数: 213 | 3 You are constantly using DH5alpha?
I am trying to clone genes into several plasmids for further transferring
into baculovirus system. Basically it is MultiBac system I am dealing with.
What I am not clear is why TOP10 was suggested not the more popular DH5alpha.
【在 s********r 的大作中提到】
: dont know your case. But we use that constantly for most of the molecular
: biology work, including large targeting vectors. They work perfectly.
|
s********r
发帖数: 312 | 4 Checked with the technician. We use "Subcloning Efficiency DH5α Competent
Cells" from invitrogen. I think TOP10 and DH5α are similar, except the
former having better efficiency.
" |
d***y
发帖数: 8536 | 5 你要做多大的？ top10 我整过1万5的质粒，毫无问题。 top10和5alpha差不多的，就
是top10的效率更高些。 |
C*******e
发帖数: 4348 | 6 1万5的质粒是啥？
1万5个constructs？
15000bp？（15 kb）？
【在 d***y 的大作中提到】
: 你要做多大的？ top10 我整过1万5的质粒，毫无问题。 top10和5alpha差不多的，就
: 是top10的效率更高些。
|
C*********m
发帖数: 213 | 7 质粒本身7.5kb左右 加两三个1000a.a.的基因总长度超过15kb-20kb
phenotype层次说top10有mcr突变，转甲基化的DNA容易一些。DH5alpha没有看到。
【在 d***y 的大作中提到】
: 你要做多大的？ top10 我整过1万5的质粒，毫无问题。 top10和5alpha差不多的，就
: 是top10的效率更高些。
|
