TA Cloning® Kits
TA Cloning® technology greatly simplifies traditional restriction and ligation cloning with a one-step cloning strategy that eliminates the need for any enzymatic modifications of the PCR product and not require the use of primers that contain restriction enzyme sites.
TA cloning® kits rely on the complementary bases of adenine (A) and thymine (T) on different DNA fragments to hybridize together. PCR products are amplified using a Taq DNA polymerase which adds a single deoxyadenosine to the 3' end of the product. The linearized vectors supplied in TA cloning® kits have a complimentary 3´ deoxythymidine (T) residues allowing the insert to ligate into the vector efficiently.
Speed with Flexibility
|With the addition of the ExpressLink™ T4 DNA Ligase into TA Cloning® Kits, ligation can now be performed at room temperature in only 15 minutes with reactions typically yielding >80% recombinants containing inserts.
TA Cloning® kits are available with a choice of pCR® 2.1 and pCR® II vectors. The T7 and Sp6 promoters of the pCR™ II vector allow in vitro transcription of the insert to produce sense or anti-sense products.
Performance and Value
TA Cloning® kits are available without competent cells (K2020-20 and K2020-40) or with One Shot® INVαF’ Chemically Competent E. coli (K2000-01 and K2000-40), and One Shot® TOP10F’ Chemically Competent E. coli (K2040-01 and K2040-40) in both 20 and 40-reaction pack sizes.
For TA Cloning® with the pCR® II Vector (dual promoter) kits are available without competent cells (K2750-20 and K2750-40), with One Shot® INVαF’ Chemically Competent E. coli (K2050-01 and K2050-40), and One Shot® TOP10F’ Chemically Competent E. coli (K2060-01 and K2060-40) in both 20 and 40-reaction pack sizes.
TA Cloning® Kits outperform ‘Competitor P’ in performance and value.
|TA Cloning® Kits||Competitor P|
|Cloning efficiency||80% (with control)||
60% (with control)
|Time for ligation||15 minutes||1 hour|
|Temperature for ligation||Room Temperature||Room Temperature|
|PCR Clean-up required?||No||Recommended|
|Selection Antibiotic||Kanamycin & Ampicillin||Ampicillin Only|
|Control Insert DNA|
|Control DNA template|
|Control PCR Primers|
Using a proofreading enzyme?
Thermostable polymerases containing extensive 3´ to 5´ exonuclease activity, such as Platinum® Pfx, do not leave 3´ A-overhangs. PCR products generated with Taq polymerase have a high efficiency of cloning in the TA Cloning® system because the 3´ A-overhangs are not removed. However, if you use a proofreading polymerase or wish to clone blunt-ended fragments, you can add 3´ A-overhangs by incubating with Taq at the end of your cycling program.
Alternatively, you may want to try the Zero Blunt® PCR Cloning Kit. This kit offers efficient cloning of blunt-end PCR products generated using thermostable, proofreading polymerases.
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