TA & Blunt End Cloning
PCR products amplified with a non-proof reading DNA polymerase such as Taq characteristically have single Adenine (A) overhangs at the 3’-end of each synthesized strand of DNA. The PCR product can then be ligated into any linearized vector having complementary single thymidine (T) nucleotides. T-vectors can be generated through the addition of a single thymidine residue to vector DNA predigested with a restriction enzyme that produces blunt ends. Linearized blunt-ended vector is treated with terminal transferase and dideoxythymidine(ddT) –use of ddT limits the reaction to the addition of a single T nucleotide.
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Which TA or Blunt End Cloning Kit is Right for You?
| General subcloning | In vitro transcription, sequencing | Blunt-end, high-efficency & less screening | |||
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| Cloning Efficiency | Up to 80% | Up to 80% | Up to 95% | ||
| Insert Sizes for Maximum Cloning Efficiency | 400-700bp | 400-700bp | 800 bp | ||
| Cloning Method | TA (Taq-Amplified inserts) | TA (Taq-Amplified inserts) | Blunt | ||
| Compatible Enzymes | Platinum Taq DNA Polymerase Platinum Taq DNA Polymerase High-Fidelity Taq Polymerase, Recombinant |
Platinum Taq DNA Polymerase Platinum Taq DNA Polymerase High-Fidelity Taq Polymerase, Recombinant |
Platinum® Pfx DNA Polymerase AccuPrime™ Pfx DNA Polymerase |
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| Positive Selection Using ccdB Gene |  |
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| Promoters | T7 | T7 & SP6 | T7 | ||
| Vector Name | pCR® 2.1 | pCR® II | pCR®-Blunt | ||
| Available One Shot® Comp Cell Options | TOP10, TOP10F', INVaF' | TOP10, INVaF' | TOP10 | ||
| Price/Reaction | $11.55 - $18.25 | $11.55 - $19.28 | $10.65 - $20.20 | ||
| Order Now | Order Now | Order Now |
