reverse PCR and blunt-end ligation

Pamela Norton pamela.norton at mail.tju.edu
Thu Nov 16 12:54:41 EST 2000


In article <Pine.GSO.4.20.0011131323050.561-100000 at hermes.ulaval.ca>,
MOUNIR IZALLALEN <aah313 at agora.ulaval.ca> wrote:

> Hi,
> 
> in order to delete a gene cloned in pbluesript, I used a pair of outward
> primers to amplify upstream and downstream the gene, and also to keep
> the
> pbluescript vector. The digestion of the PCR  analysis gives the results
> anticipated. But The selfligation of that product failed, and I tried
> many concentrations with no success. So I'm wondering, If I have to kinase
> the PCR product prior to ligation.
> 
> Thanks

In case no one has answered your question, yes, you must either kinase
the PCR product or the starting primers. 

Pam Norton






More information about the Methods mailing list