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.
This promotion is open to customers in the US and Canada. Discount will apply to qualifying orders received by Life Technologies no later than July 31, 2013 or until promotional supplies are depleted, whichever comes first. Discount applies to list price in effect at the time order is received by Life Technologies. Cannot be combined with other discounts or promotions. Offer void where prohibited, licensed or restricted by federal, state, provincial, or local laws or regulation or agency/institutional policy. Other restrictions may apply.
For research use only. Not for use in diagnostic procedures.