Overview: TA Cloning exploits the terminal transferase activity of some DNA polymerases such as Taq polymerase. This enzyme adds a single, 3'-A overhang to each end of the PCR product. This makes it possible to clone this PCR product directly into a linearized cloning vector with single, 3'-T overhangs. The PCR products with dA overhang, are mixed with this vector in high proportion. The complementary overhangs of "T" vector and PCR product will be ligated under the action of T4 DNA ligase.
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Protocols
- TOPO TA Cloning (Linder Lab, University of Texas)
Describes ligation reaction, transformation and clone selection.
http://www.esb.utexas.edu/anneke/LinderLab/protoco...
Added: Tue Apr 13 2004, Hits: 4998, Reviews: 0 Write review Cached - Addition of 3'-A Overhangs (A-Tailing) to PCR Product (Protocol Online)
PCR product amplified by proofreading polymerases is blunt-ended because proofreading polymerases possess 3´→5´ exonuclease activity that removes the 3´-A overhangs necessary for TA cloning. However, 3´-A overhangs can be added to blunt-end fragments using a Taq polymerase after PCR amplification.
Added: Mon Jan 02 2006, Reviews: 0 Write review -
Construction of Homemade 'T-vectors'
(Dr. Simon Dawson, Department of Biochemistry, University of Nottingham Medical School)
Making T vector from plasmid for TA cloning
http://www.nottingham.ac.uk/~mbzspd/methods/Constr...
Added: Sat Apr 10 2004, Hits: 3000, Reviews: 0 Write review Cached -
T/A Cloning
(Practical Approach Online, Oxford University Press)
T/A-based subcloning of PCR amplified DNA fragment
http://www.oup.co.uk/pdf/pas/9v1-7-14.pdf
Added: Sat Aug 17 2002, Hits: 8998, Reviews: 0 Write review -
TA Cloning and Vector Preparation
(Janos Luka Ph.D., Department of Pathology, Eastern Virginia Medical School)
It is relatively easy and cheap to make a vector containing a T overhang. Because Taq polymerase ads an A overhang to PCR products in standard PCR assays, almost any PCR product can be cloned into this vector. The purified T-vector can be stored aliquoted at -70 C for at least one year. This protocol describes how to make T vectors and how to clone PCR products.
http://herpesvirus.tripod.com/research/T-vectorclo...
Added: Fri Apr 16 2004, Hits: 6911, Reviews: 0 Write review Cached -
TA Subcloning of PCR Products
(Gerard Lazo)
http://wheat.pw.usda.gov/~lazo/methods/lazo/met7.h...
Added: Tue May 14 2002, Hits: 4995, Reviews: 0 Write review Cached