Description:
OneTaq® Hot Start 2X Master Mix with GC Buffer is an optimized blend of Taq and Deep VentR ™ DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to PCR applications from GC-rich templates including pure DNA solutions, bacterial colonies and cDNA products. The 3´→5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase (1). The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient master mix formulation contains dNTPs, MgSO4 and other buffer components and stabilizers listed below, requiring only the addition of primers and DNA template for robust amplification.
Source:
An E. coli strain that carries the Taq DNA Polymerase gene from Thermus aquaticus YT-1 and an E. coli strain that carries the Deep VentR DNA Polymerase gene from Pyrococcus species GB-D. Applications:
GC-rich PCR
High Sensitivity PCR
High Throughput PCR
Colony PCR
Long PCR (up to ~6 kb genomic)
Hot Start PCR
Reagents Supplied:
OneTaq High GC Enhancer
OneTaq Hot Start Master Mix with GC Buffer
Heat Inactivation:
No
Reaction Conditions:
OneTaq Hot Start Master Mix with GC Buffer OneTaqHot Start Master Mix with GC Buffer:
80 mM Tris-SO4
20 mM (NH4)2SO4
0.2 mM dNTPs
2 mM MgSO4
5 % Glycerol
0.05 % Tween-20
5 % DMSO
0.06 % IGEPAL® CA-630
25 units/ml OneTaq Hot Start DNA Polymerase
pH 9.2 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 15 nmol of dNTP into acid insoluble material in 30 minutes at 75°C. Unit Assay Conditions:
1X ThermoPol® Reaction Buffer, 200 μM dNTPs including [3H]-dTTP and 200 μg/ml activated Calf Thymus DNA. Reaction Conditions:
1X OneTaq Hot Start Master Mix with GC Buffer, DNA template and primers in a
total reaction volume of 25–50 μl. OneTaqHigh GC Enhancer:
10 mM Tris-HCl
25% DMSO
25% Glycerol
pH 9.2 @ 25°C Concentration:
2 X
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