PCR after T4 ligation? - (Jul/30/2014 )
Greetings friends and colleagues, I'm just wondering, if It's possible to make a pcr after a t4 ligation, without preivious EtOH precipitation? I mean... The T4 buffer could be a problem for the PCR reaction?
Thanks for read!
You can use vanishingly small amounts of your ligation product in your PCR reaction, so the effect will be near zero.
Well Amaizing! That's sounds obvious, but I had to be sure, thanks a lot!
I've done this before. I used 1uL of my ligation in a 50uL PCR reaction (using NEB's Phusion polymerase and HF buffer) and it worked great for me.