TA Cloning Vectors
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.
More information about the Methods