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.
incubate with PFU polymerase and dNTP for 30 mins at 72oC
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.