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.

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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 efficiency80% (with control)

60% (with control)

Time for ligation15 minutes1 hour
Temperature for ligationRoom TemperatureRoom Temperature
PCR Clean-up required?NoRecommended
Selection AntibioticKanamycin & AmpicillinAmpicillin Only
 

Kit Contents

Vector 
Ligation Buffer 
Control Insert DNA  
dNTPs 
Sterile Water 
Control DNA template 
PCR Buffer 
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.

For research use only. Not for use in diagnostic procedures.