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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. blink.gif

-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-