2x Pfu PCR Master Mix
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.
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
|Template DNA(<1μg)||< 1 μl||as required|
|Forward Primer(20 μM)||1 μl||0.2-0.4 μM each|
|Reverse Primer (20 μM)||1 μl||0.2-0.4 μM each|
|2xPfu PCR master Mix||25 μl||1x|
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
|PCM11||2x Pfu PCR Super Master Mix||Pfu PCR Mix, without dye. 1ml/vial or bulk|
|PCM11B||2x Pfu PCR Super Master Mix, with blue dye||Pfu PCR Mix, with dye. 1ml/vial or bulk|