make PCR template blunt ended - (Mar/01/2007 )
I want to make a long template PCR blunt ended for blunt end cloning. I have amplified the 6.4 kb PCr with Roche long template PCR system. Can anybody tell me how to make the PCR product blunt ended.
-sktewary-
incubate with PFU polymerase and dNTP for 30 mins at 72oC
-adrians-
your ends are probably already blunt ended. Proof reading enzymes more or less produce only blunt end products.
A tailing only happens with taq polymerase.
-perneseblue-