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2x Pfu PCR Master Mix

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2x Pfu PCR Master Mix

Description

It is a ready-to-use reagents containing Pfu DNA Polymerase , dNTPs, MgCl2 and PCR reaction Buffer in one PCR tube. Pfu master Mix provides robust PCR performance in a wide range of PCR applications without time-consuming optimization. Especially, this enzyme has a high PCR fidelity with improved yield and specificity. To prepare the final PCR, only primers and template DNA are added. Pfu master Mix contributes to highly reproducible PCR by reducing the risk of pipetting errors and miscalculation.

Feature

  • Premixed ready-to-use.
  • 3′->5′ proofreading activity
  • 5′->3′ exonuclease activity
  • Extra A addition (terminal transferase) activity

Applications

  • High Fidelity PCR experiments
  • Blunt End Cloning
  • Sequencing
  • Site-Directed Mutagenesis

Marks

1.Pfu Polymerase has the 3′->5′ exonuclease activity, the extend speed is lower than Taq, should increase it’s extending time according the length of template. If Template< 4 KB, set the extending speed to 1KB/min and if template>4 KB, 0.5KB/min will be better.

2.Pfu amplification need the high purity primers, and the primer should length than 18 bases, and Tm between 55-80°C.

3.Pfu Polymerase is thermal stable, with the high GC rich template the denaturation temperature can up to 98°C, which do not affect the Pfu activity.

Reaction Mixture Set Up

ComponentsVolume FinalConcentration
Template DNA(<1μg)< 1 μlas required
Forward Primer(20 μM)1 μl0.2-0.4 μM each
Reverse Primer (20 μM)1 μl0.2-0.4 μM each
2xPfu PCR master Mix25 μl1x
ddH2O22 μl 

PCR cycle

94°C 2-5 min
94°C 30 sec
55 °C 30 sec, 30-35 cycles
72°C 0.5 kb/min
72°C 5-10 min

Order

PCM112x Pfu PCR Super Master MixPfu PCR Mix, without dye. 1ml/vial or bulk
PCM11B2x Pfu PCR Super Master Mix, with blue dyePfu PCR Mix, with dye. 1ml/vial or bulk

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