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LA DNA Polymerases Mix

Description

LA DNA polymerases Mix is unique blend of two thermostable DNA polymerases. One of them is Taq DNA polymerase which is highly processive but lacks a 3’->5’ exonuclease proofreading activity, which accounts for relatively high error rate during DNA amplification. Second polymerase is less processive but possesses 3’->5’ exonuclease activity necessary for proofreading. The polymerases mix decreases error rate during synthesis of complementary DNA strands and preserves high speed of DNA synthesis. The repair capability allows the polymerases to resume elongation of the growing DNA strand and amplification of complex genomic DNA fragments up to 20 kbps. Using less complex templates, such as bacterial genomic DNA or viral DNA, amplifications of up to 20 kbps or 40 kbps, respectively, have been achieved. The fidelity of LA DNA Polymerases Mix is  approximately 5 times greater than Taq DNA polymerase. The polymerases mix is intended for "Long and Accurate" (LA) PCR. It is especially suitable for amplification of DNA fragments designated for cloning purposes. Importantly, the polymerases blend creates DNA amplicons with mostly blunt ends as a result of the proofreading activity of the polymerases.

Technical data

Components and packaging

  • LA DNA polymerases Mix is supplied at a concentration 5 U/µl. Basic packaging is 1 tube with 500 U/100 µl (Cat. No. L072), 5 tubes with 500 U/100 µl (L073) or 10 tubes with 500 U/100 µl (L074).
  • Each test tube of LA DNA polymerases Mix is accompanied with a test tube with 10x concentrated LA PCR reaction buffer containing MgCl2 (1.5 ml). If different concentration of  MgCl2 is required, a test tube with 10x concentrated LA PCR reaction buffer without MgCl2 (1.5 ml) and a separate test tube with 25 mM MgCl2 (0.5 ml) should be ordered (Cat. No. L076).
  • Each package of LA DNA polymerases mix also contains 1 test tube with DMSO (1 ml).

Storage

  • At temperature -20oC ± 5°C. Material can be repeatedly defrosted.

Composition

  • Storage buffer for LA DNA polymerases Mix: 20 mM Tris-HCl, pH 8.0 (25oC), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Nonidet P-40, 0.5% Tween 20, 50% glycerol.
  • 10x LA PCR reaction buffer: 500 mM Tris-HCl (pH 9.3 at 25oC), 150 mM (NH4)2SO4, 1% Tween 20, 22.5 mM MgCl2.

Activity

  • One unit is defined as the amount of enzyme, which catalyzes incorporation of 10 nmol dNTPs within 30 min at 72oC into trichloracetic acid precipitable material. Reaction conditions are as follow: 10 mM Tris-HCl (pH 8.8 at 25oC), 50 mM KCl, 0.1% Triton X-100, 1.5 mM MgCl2, 200 µM dATP, dCTP, dGTP and [α-32P]dTTP, 50 µg/ml denatured cDNA, 0.5 µM primer and 0.2 – 0.5 U of enzyme in the volume of 50 µl.

Purity and quality control

  • Purity of the polymerases is tested by SDS-PAGE. Material is free of nucleases.
  • Each batch of LA DNA polymerases Mix is tested for its ability to amplify DNA fragment from mammalian genomic DNA by PCR. The results are verified by electrophoresis in agarose gel in the presence of ethidium bromide; only DNA band of the expected size is present.

Downloads

Cat. No. Product name and specification Amount Price
L072 LA DNA Polymerases Mix 500 U 116.25 EUR
L073 LA DNA Polymerases Mix 5x 500 U 465 EUR
L074 LA DNA Polymerases Mix 10x 500 U 813.75 EUR
L072a LA DNA Polymerases Mix + PCR dNTP mix 500 U + 100 µl 132.5 EUR
L073a LA DNA Polymerases Mix + PCR dNTP mix 5x 500 U + 5x 100 µl 530 EUR
L074a LA DNA Polymerases Mix + PCR dNTP mix 10x 500 U + 10x 100 µl 927.5 EUR