b******r
发帖数: 111 | 1 I am cloning 3.4kb human genome DNA into a 8.4kb vector.Isolate some
plasmids. Recut,then 3.4kb bands
appear. But sequencing shows it is not 3.4kb human DNA,it is a unknown
fragment. Strangely,the size is same
as 3.4kb! What happen? Any idea to eliminate recombination ? The competent
cells used are Invitrogen ElectroMAX™ DH10B™ T1 Phage-
Resistant Competent Cells Cat. No. 12033-015. | Z******5
发帖数: 435 | 2 把链接之前的PCR产物测序一下，看看对不对。
此外，在目的基因内部找一个或两个常用的酶切位点，在PCR之后和克隆测序之前酶切
看看条带大小对不对，也可以作为初步判断的依据。 | M*****n
发帖数: 16729 | 3 if you are concerned about recombination, use Stbl4 cells. |
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