- Robust amplification of long DNA templates (long range PCR) – obtaining products up to 48 kb is possible with minimal reaction optimization
- Accurate, long PCR amplification of genomic targets
- DNA proofreading activity allows for high fidelity amplification from a variety of templates
- High yields are possible with the robust enzyme-buffer system – includes an optimized buffer (LA Buffer II)
|TaKaRa LA Taq DNA Polymerase ||250 U (5 U/µL)*|
|10X LA PCR Buffer II (contains 25 mM MgCl2) ||1 mL|
|dNTP Mixture (2.5 mM each dNTP) ||800 µL|
| || |
|TaKaRa LA Taq DNA Polymerase ||125 U (5 U/µL)*|
|10X LA PCR Buffer II (without Mg2+) ||1 mL|
|25 mM MgCl2 ||1 mL|
|dNTP Mixture (2.5 mM each dNTP) ||400 µL|
|*Protocol recommends the use of 2.5 U per 50 µL reaction.|
Avoid repeatedly freezing and thawing TaKaRa LA Taq and dNTPs. Once thawed, aliquot into separate tubes and store at –20°C.
PCR products generated with LA Taq contain a mixture of 3'-A overhangs and blunt ends which allows >80% cloning efficiency into T-vectors. However, the efficiency of cloning longer products (>5 kb) into T-vectors is quite low.
Enzyme performance is confirmed by successful amplification of a 35 kb lambda DNA template and a 17.5 kb human genomic DNA template.