Subcloning large PCR products

Paul E. Ulanch PULANCH at bioch.tamu.edu
Tue Jan 30 17:58:04 EST 1996


In article <1996Jan29.194614.8132 at mcrcr6>, 
rosasg01 at mcrcr6.med.nyu.edu wrote:
>hElP!!!
>
>For the last 3 months I have tryed to subclone a 8kb PCR product. 
So
>far, nothing has worked. Does somebody has done something like 
this???
>I'm sure somebody must have done it. If you have successfully
>subcloned large PCR products, please let me know how big it was and
>the protocol that you used for both PCR and subcloning. I'll really
>appreciate any suggestions.
>Thanks.
>German Rosas-Acosta, graduate student
>NYU Med. Center
>I

What enzyme are you using in the PCR reaction?  Is it Taq or a 
mixture of enzymes (hint: 3' to 5' exonuclease present?)

Paul "Goose" Ulanch

Dept. Biochem/Biophys
Texas A&M University
College Station, TX   77843-2128
(409) 847-9379
PULANCH at bioch.tamu.edu



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