olligo annealing for shRNA cloning - (May/25/2009 )
In the protocol you mentioned above,
5ul Forward oligo
5ul Reverse oligo
5ul 10x NEB buffer2
Is the "10X NEB buffer2" same as the reaction buffer they provide with restriction enzyme?
I have the exactly same problem now, and in my case, I am using 10x annealing buffer from Gendepot.
After several times of failure, I am not a fan of their annealing buffer. I am up to follow new protocol.