Description:
OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer is an optimized, ready-to-use blend of Taq and Deep VentR™ DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to routine PCR applications from 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 TaqDNA Polymerase (1). The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient quick-load master mix formulation contains dNTPs, MgCl2, buffer components and stabilizers as well as two commonly used tracking dyes for DNA gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~4 kb and Tartrazine migrates at ~10 bp. Both dyes are present in concentrations that do not mask co-migrating DNA bands.
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:
High Sensitivity PCR
High Throughput PCR
Routine PCR
AT-rich PCR
Colony PCR
Long PCR (up to ~6 kb genomic)
Hot Start PCR
Reagents Supplied:
OneTaq Hot Start Quick-Load Master Mix with Standard Buffer Heat Inactivation:
No
Reaction Conditions:
1X OneTaq Hot Start Quick-Load Master Mix with Standard Buffer 1X OneTaqHot Start Quick-Load Master Mix with Standard Buffer:
20 mM Tris-HCl
Xylene Cyanol FF
22 mM KCl
0.2 mM dNTPs
1.8 mM MgCl2
5 % Glycerol
0.05 mM Tween-20
0.06 % IGEPAL® CA-630
22 mM NH4Cl
Tartrazine
25 units/ml OneTaq Hot Start DNA Polymerase
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 Quick-Load Master Mix with Standard Buffer, DNA template and primers in a total reaction volume of 50 µl. Concentration:
2 X