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!
-jesscalleros-
You can use vanishingly small amounts of your ligation product in your PCR reaction, so the effect will be near zero.
-phage434-
Well Amaizing! That's sounds obvious, but I had to be sure, thanks a lot!
-jesscalleros-
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.
-Bio-Lad-