pfu for cloning PCR

Franck Laurent franck.laurent at medew.fyto.wau.nl
Tue May 6 06:44:56 EST 1997


In article <3367C9D9.7F7D at psuvm.psu.edu>, zxz4 at PSUVM.PSU.EDU says...
>
>>
>> 
>> 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.


If the PCR does'nt work be carefull with the extension time. Pfu works much 
more slowly than many polymerases (2 min/kb)

Franck





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