reverse PCR and blunt-end ligation

MOUNIR IZALLALEN aah313 at agora.ulaval.ca
Mon Nov 13 13:31:56 EST 2000


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







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