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OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer
OneTaq DNA Polymerase is an optimized blend of Taq and Deep Vent® DNA polymerases for use with routine and difficult PCR experiments.
• Unique hot start formulation eliminates nonspecific amplification and does not require a separate activation step
• Obtain high yields with GC rich templates (>65%)
• Offered with inert tracking dyes for easy and direct loading of PCR products onto gels
产品信息
OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer is an optimized, ready-to-use blend of Taq and Deep Vent® 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, 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.
重点
产品来源
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 Vent® DNA Polymerase gene from Pyrococcus species GB-D.
产品类别:
OneTaq® DNA Polymerases Products,
Taq DNA Polymerase Products,
Master Mixes Products
应用:
Hot Start PCR,
Multiplex PCR,
Specialty PCR,
Routine PCR,
PCR
产品组分信息
产品组分信息
本产品提供以下试剂或组分:
NEB #
名称
组分货号
储存温度
数量
浓度
M0489S
-20
OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer
M0489SVIAL
-20
2 x 1.25 ml
2 X
OneTaq® High GC Enhancer
B9026AVIAL
-20
1 x 2 ml
不适用
M0489L
-20
OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer
M0489SVIAL
-20
10 x 1.25 ml
2 X
OneTaq® High GC Enhancer
B9026AVIAL
-20
3 x 2 ml
不适用
特性和用法
1X 预混液组分
80 mM Tris-SO4 1X Tartrazine 25 units/ml OneTaq Hot Start DNA Polymerase 20 mM (NH4)2SO4 0.2 mM dNTPs 2 mM MgSO4 5% Glycerol 5% DMSO 0.06% IGEPAL® CA-630 0.05% Tween® 20 1X Xylene Cyanol pH 9.2 @ 25°C
热失活
否
单位活性检测条件
1X ThermoPol® Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and 15 nM primed M13 DNA.
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.
Reaction Conditions: 1X OneTaq Hot Start Quick-Load Master Mix with GC Buffer, DNA template and primers in a total reaction volume of 50 µl.
优势和特性
应用特性
GC-rich PCR
High Sensitivity PCR
High Throughput PCR
Colony PCR
Long PCR (up to ~6 kb genomic)
相关产品
相关产品
Magnesium Sulfate (MgSO4) Solution
OneTaq® DNA Polymerase
OneTaq® Hot Start 2X Master Mix with GC Buffer
OneTaq® Hot Start 2X Master Mix with Standard Buffer
OneTaq® Hot Start DNA Polymerase
OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer
注意事项
Product specifications for individual components in the OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer are available separately.
OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer is stable for fifteen freeze-thaw cycles when stored at -20°C.
OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer is also stable for one month at 4°C, so for frequent use, an aliquot may be kept at 4°C.
参考文献
Barnes, W.M. (1994). Proc. Natl. Acad. Sci. USA. 91, 2216-2220.
Saiki R.K. et al. (1985). Science. 230, 1350-1354.
Powell, L.M. et al. (1987). Cell. 50, 831-840.
操作说明、说明书 & 用法
操作说明
Protocol for OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer
Guidelines for PCR Optimization with OneTaq® and OneTaq® Hot Start DNA Polymerases
使用指南
Activity of Restriction Enzymes in PCR Buffers
Guidelines for PCR Optimization with OneTaq® and OneTaq® Hot Start DNA Polymerases
应用实例
Robust Colony PCR from Multiple E. coli Strains using OneTaq® Quick-Load® Master Mixes
工具 & 资源
Web 工具
Tm Calculator
FAQs & 问题解决指南
FAQs
How should I set up a PCR using the OneTaq® Hot Start Master Mixes?
Can I use my regular Taq-based cycling conditions for OneTaq® Hot Start DNA Polymerase based products?
How do I activate OneTaq® Hot Start Polymerase?
What are the stability and storage requirements of the Quick-Load® OneTaq® Master Mixes?
What type of DNA ends result from a primer extension reaction or a PCR using OneTaq® DNA Polymerase?
How long a product can be made by OneTaq® DNA Polymerase?
Can OneTaq® DNA Polymerase be used with uracil-containing primers or bisulfite-treated DNA?
What is the fidelity of OneTaq® DNA Polymerase?
When should I add the High GC Enhancer?
Where can I find help troubleshooting my PCR?
How should I determine an appropriate annealing temperature for my reaction?
How is OneTaq DNA Polymerase different from LongAmp™ Taq DNA Polymerase?
实验技巧
A master mix with a gel loading dye is the ultimate time-saver! It allows for easy reaction set up and with the gel loading dye already in the mix, the results of your PCR are available even faster.
引用 & 技术文献
引用文献
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专题文章
Using aptamers to control enzyme activities: Hot Start Taq and beyond