Re: help: Molecualr cloning problem# Biology - 生物学
g*m
1 楼
check NEB catalog for Taq enzyme, it has double strand specific 5'->3'
exonuclease activity.
My guess is primer and/or product degradation by Taq.
BTW, one small tip for pfx is: using 2X buffer rather than 1X. I got several
cDNA clones by PCR, and none of them disrupt the ORF by this modification.
DNA(WASP)
然后用同样的primer
colony-pGEX4t1
这些在处理后的pGEX4t1
exonuclease activity.
My guess is primer and/or product degradation by Taq.
BTW, one small tip for pfx is: using 2X buffer rather than 1X. I got several
cDNA clones by PCR, and none of them disrupt the ORF by this modification.
DNA(WASP)
然后用同样的primer
colony-pGEX4t1
这些在处理后的pGEX4t1