• Reduced Background
• Increased Specificity
• High Performance

Description

TaKaRa LA Taq™ HS is designed to be suitable for Hot Start PCR. It is derived from TaKaRa LA Taq™ and neutralizing monoclonal antibody to Taq DNA polymerase. Non-specific amplification due to mispriming and/or formation of primer dimer before thermal cycling can be prevented, since the antibody inhibits the polymerase activity by binding to the Taq DNA polymerase until the temperature elevates. This enzyme can be also used in general PCR conditions, since monoclonal antibody is denatured in the initial DNA-denaturation step.

RR042A/B
*Supplied with 10 × LA PCR Buffer II (Mg²⁺ plus) and dNTP Mixture

Storage

-20°C

Concentration

5 units/µl

Form

20 mM Tris-HCl (pH8.0), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20, 0.5% Nonidet P-40, 50% Glycerol

Purity

Nicking activity, endonuclease and exonuclease activity are not detected after the incubation of 0.6 µg of supercoiled pBR322 DNA, 0.6 µg of λDNA or 0.6 µg of λ-Hind III digest with 10 units of this enzyme for 1 hour at 74°C.

PCR performance test

Good performance of DNA amplification by PCR is confirmed by using λDNA as the template (amplified fragment: 35 kbp).
Good performance of DNA amplification of β-globin gene by PCR is also confirmed by using human genomic DNA as the template (amplified fragment: 17.5 kbp).
Inhibition of LA Taq activity by the antibody is confirmed to be more than 90% after the reaction at 55°C for 10 min.

PCR products

As most PCR products amplified with TaKaRa LA Taq™ HS have one A added at 3′-termini, the obtained PCR product can be directly used for cloning into T-vector. Also it is possible to clone the product in blunt-end vectors after blunting and phosphorylation of the end.

10 × LA PCR Buffer II (Mg²⁺ plus) supplied

Volume:	1 ml/vial
Mg2+ concentration (10 ×):	25 mM

dNTP Mixture supplied

Mixture of dNTP, ready for use in PCR without dilution

Volume:	400 µl/vial
Concentration:	2.5 mM of each dNTP
pH:	7-9
Form:	Solved in water (sodium salts)
Purity:	≥98% for each dNTP

General reaction mixture for PCR (total 50 µl)

TaKaRa LA Taq™ HS (5 units/µl)	0.5 µl
10 × LA PCR Buffer II (Mg2+ plus)	5 µl
dNTP Mixture (2.5 mM each)	8 µl
Template	< 1 µg
Primer 1	0.2-1.0 µM
Primer 2	0.2-1.0 µM
Sterilized distilled water	up to 50 µl