How to deal with so many false positive clones - Transfer linear DNA into E.coli competent cells... (Nov/14/2008 )

Hello,

I have a problem in using Red recombination system to knockdown bacteria genes. I transfered linear DNA (long homologous arms) into E.coli competent cells by electroporation. But when I tested its sensitivity with kanamycin, there are so many false positive clones on the plate but no positive ones. How can I do?

Thanks,
Yuri

-yurippe-

QUOTE (yurippe @ Nov 14 2008, 05:08 AM)

You need two arms of homology, otherwise you will get many false integrations. Also change the length or placement of homologous regions, if this continues to be a problem.

-cellcounter-

thanks very much! I will try again.

-yurippe-