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Pfu DNA Polymerase Ⅱ

DNA polymerase、 DNA polymerase B、Pfu polymerase、Pol I

价格￥820.00 （供应商现货： 3-5个工作日）

货号 UA070072

规格

250U1000U

数量

产品介绍

产品介绍评论(0)

产品规格

分子别名

DNA polymerase、 DNA polymerase B、Pfu polymerase、Pol I
表达宿主

E.coli
分子量

90 kDa (Reducing)
纯度

＞95% by SDS-PAGE
活性

5 U/μl
标签

His Tag
性状

Liquid
缓冲体系

20 mM Tris-HCl, 0.1 mM EDTA, 0.1% Tween20, 0.1% triton X100, 1 mM DTT, 100 mM KCl, 50%Glycerol、pH 8.2 @ 25°C
储存条件

Store at -25 ~ -15℃ for 2 years
文献引用

[1] Lin, T. C. , et al. "Cloning and expression of T4 DNA polymerase." Proceedings of the National Academy of Sciences 84.20(1987):7000-7004.
[2] Yan, Wang , et al. "A novel strategy to engineer DNA polymerases for enhanced processivity and improved performance in vitro." Nucleic Acids Research 3(2004):1197.

背景介绍

Pfu DNA Polymerase is a thermostable enzyme that replicates DNA at 75°C. It catalyzes the polymerization of nucleotides into duplex DNA in the 5´→3´ direction in the presence of magnesium. The enzyme has a molecular weight of approximately 90,000 daltons as estimated from the predicted amino acid sequence and exhibits 3´→5´ exonuclease (proofreading) activity. Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high fidelity.

产品组分

Storage Solution : 5 U/ul Pfu DNA Polymerase Ⅱ, 20 mM Tris-HCl, 0.1 mM EDTA, 0.1% Tween20, 0.1% triton X100, 1 mM DTT, 100 mM KCl, 50%Glycerol、pH 8.2 @ 25°C
10* Reaction Buffer: 200 mM Tris-HCl (pH 8.8) , 20 mM MgSO4, 100 mM KCl, 100 mM (NH4) 2SO4, 1% Triton X-100, 1 mg/ml nuclease-free BSA

操作步骤

1. Set up a 50 μl PCR reaction system as follows:

2. Recommended thermal cycling conditions for Pfu DNA Polymerase-mediated PCR amplification as follows:

Step	Temperature	Time	Number of Cycles
Initial Denaturation	95°C	1–2 minutes	1 cycle
Denaturation Annealing* Extension	95°C 42–65°C 72–74°C	0.5–1 minute 30 seconds 2–4 minutes	25–35 cycles
Final Extension	72–74°C	5 minutes	1 cycle
Soak	4°C	Indefinite	1 cycle

*The annealing temperature for a specific amplification reaction will depend upon the sequences of the two primers.

注意事项

Note: It is critical to withhold Pfu DNA Polymerase until after the addition of dNTPs; otherwise, the proofreading activity of the polymerase may degrade the primers,resulting in nonspecific amplification and reduced product yield. Assemble on ice.

酶活定义

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C