b******n
发帖数: 4225 | 1 呵呵，抽质粒老手也碰上这种奇怪的事情
某种Gram-菌的一个isolate/strain，通过conjugation转进了shuttle vector之后
抽质粒用的是Qiagen mini-prep kit
跑电泳发现是很亮的smear
作为对照，该种菌的另外一个isolate/strain同时进行了conjugation和plasmid
extraction得到了清晰的质粒超螺旋条带
做了几次都是这样
现在有点迷糊，不知道可能的问题所在
谢谢指点 |
h*****9
发帖数: 4028 | 2 that strain/isolate has endonuclease activity? wash with PB buffer before
using PE. |
b******n
发帖数: 4225 | 3 yes, I washed once
according to reports, this isolate/strain has strong endonuclease activity
then any suggestion?
【在 h*****9 的大作中提到】
: that strain/isolate has endonuclease activity? wash with PB buffer before
: using PE.
|
h*****9
发帖数: 4028 | 4 Wash one more time, eluted DNA using 1xTE
【在 b******n 的大作中提到】
: yes, I washed once
: according to reports, this isolate/strain has strong endonuclease activity
: then any suggestion?
|
