Pfu DNA Polymerase
DNA polymerase、 DNA polymerase B、Pfu polymerase、Pol I
- 价格: ¥820
- 规格:
- 数量:

产品介绍
- 产品介绍
- 操作详情
产品组分
Storage Solution : 5 U/ul Pfu DNA Polymerase, 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:
Reagent
Volume
Final Concentration
10X Pfu Buffer (with Mg2+)
5µl
1 X
dNTP mix, 10mM each
1µl
200µM each
upstream primer
5–50pmol
0.1–1.0µM
downstream primer
5–50pmol
0.1–1.0µM
DNA template
variable
<0.5µg/50µl
Pfu DNA Polymerase (5U/µl)
variable
1.25U/50µl
DEPC-treated Water
Up to 50µl
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2. Recommended thermal cycling conditions for Pfu DNA Polymerase-mediated PCR amplification as follows:
*The annealing temperature for a specific amplification reaction will depend upon the sequences of the two primers.注意事项
Note: 1. 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.
2. Assemble on ice.
- 产品数据
生物活性
In the experiment, the 773bp target fragment was amplified by PCR using PUC57 as the template. At the same time, competing enzymes were added to investigate the enzyme activity of Pfu DNA Polymerase. As shown in the figure, this product has the following effects.
Lane 1 Negative Control (negative control with no added enzyme only);
Lane 2 UA-Pfu DNA Polymerase 2.5U;
Lane 3 UA-Pfu DNA Polymerase 5U;
Lane 4 Competing product A 5U;
Lane 5 Competing product B 5U;
电泳(SDS-PAGE)
1μg (R: reducing condition, N: non-reducing condition).
体积排阻色谱(SEC-HPLC)
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