Platinum™ Taq DNA Polymerase

PCR Hot start PCR - Bacterial DNA

Experiment
PCR Hot start PCR - Bacterial DNA
Product
Platinum™ Taq DNA Polymerase from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
Always thaw on ice

Publication protocol

After thawing at room temperature, the samples
were subjected to the process of DNA extraction with
the DNeasy Blood & Tissue Kit (QIAGEN, Germany).
Then, a PCR was carried out to check the efficiency
of extraction of DNA using the primer pair ISO2G14.
DNA amplification was carried out in a final volume
of 25 µL, containing 5 U of Platinum Taq DNA
Polymerase, 5 pmol of primers pair, 5 µL of reaction
buffer 5X, 25 mM of MgCl2, 10 mM of dNTP, 2 µL
of sample and water. The PCR cycling protocol was:
95° C for 2 min, 40 cycles of 95° C for 1 min, 50° C
for 1 min and 72° C for 1 min, elongation at 72° C for
5 min

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Reviews

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Discussion

Discussion

4 years ago

Author: Qatar

When should I add the polymerase for hot start PCR?

I keep getting a non-specific band in PCR so I would like to try a hot start PCR manually. How should I prepare the mix and at which step should I be adding the polymerase?

Discussion

4 years ago

Author: Janina Reinhardt Switzerland

Hot start PCR using phusion green DNA polymerase

I am currently performing hot start PCR using phusion green hot start DNA polymerase in order to amplify a 8.8kb insert. However, I cannot get any amplified product at all. I have tried changing some parameters but I am not sure what to do. Any help?

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Papers

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Platinum™ Taq DNA Polymerase below.

Download PDF Download manufacturer protocol

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