pfu for cloning PCR

[Zhiyong Zhao]

>
> 
> Subject: pfu for cloning PCR?
> 
> Date: 30 Apr 1997 17:08:55 GMT
> 
> Hi all, does anybody know if pfu can be used for cloning PCR. I know
> taq
> is suitful for it. The reason I am using pfu is that I has tons of
> pfu. It
> has tried several times by many person in our lab in different
> experiments and seemed no success. I dont know if someone has similar
> problems as I have.
> Thanks everybody, I like you.
> 
> C.Lu
> C.Lu at cellbio.duke.edu

Dr. Duncan Clark has just summarized the polymerases used in PCR (see 
"Re: Overhang A of PCR". 4/29). The PCR products generated by pfu are 
supposed to be ligated into the vector with blunt ends since the enzyme 
has proofreading activity. I do not quite get what you meant "no 
success". Is it "no success" in PCR or ligation? If your PCR works with 
pfu, the problems are in ligation.


Zhiyong Zhao, Ph.D.