TA Cloning Vectors

Tracy Aquilla aquilla at salus.med.uvm.edu
Tue Feb 21 08:03:48 EST 1995


In Article <h-petrie-1702950929530001 at mac-95.k-g2.ski.mskcc.org>,
h-petrie at ski.mskcc.org (Howard T. Petrie) wrote:
>In article <sg-1602950954500001 at pc030.surgery.nwu.edu>, sg at nwu.edu
>(Stephen Gately) wrote:
>
>> Hi, an old question to this group.  What TA cloning vector have
>> individuals used sucessfully for cloning PCR amplified cDNA fragments of
>> approximately 600-800 bp...
>
>
>
>
>We have used the InVitrogen TA cloning system a number of times with
>excellent results and no problems.  It's kind of hard to argue with that.

Well, I'll take that challenge. The assumption that every PCR reaction
results in a product with an overhanging A is a very poor one, particularly
if you use a polymerase other than Taq. TA cloning systems are generally
unreliable, due to this fact. For details, you should read DNA and Cell
Biology, vol. 12(8): 763-770. "DNA Polymerase-Catalyzed Addition of
Nontemplated Extra Nucleotides to the 3' End of a DNA Fragment", by Gengxi
Hu, 1993. This paper has a nice table, listing the terminal residues
normally found on the ends of PCR products amplified by various polymerases.
This is good reading for anyone cloning PCR products.
    tracy



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