KAPA LongRange

PCR Hot start PCR - Mammalian DNA

Experiment
PCR Hot start PCR - Mammalian DNA
Product
KAPA LongRange from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
This product can also be used to amplify mid to long-range targets (up to 15 kb)

Publication protocol

Enzymes and Amplicons

We evaluated six commercially available long-range enzymes including SequalPrep polymerase (Invitrogen, Carlsbad, CA), AccuPrime Taq DNA Polymerase (Invitrogen, Carlsbad, CA), PrimeSTAR GXL polymerase (TaKaRa Bio, Shiga, Japan), LA Taq Hot Start Version Polymerase (TaKaRa Bio, Osaka, Japan), KAPA long Range HotStart DNA polymerase (KAPA Biosystems, Wobum, MA) and QIAGEN LongRange PCR Polymerase (Hilden, Germany). These enzymes were selected based on our knowledge at the time of the experiments, and based on Internet search. However, this is not a comprehensive list, and we acknowledge that other similar enzymes are also commercially available, such as New England Biolabs Phusion HF Polymerase and LongAmp Taq DNA Polymerase, Roche Expand Long Range DNA Polymerase, etc. Readers should not assume that the six enzymes used in the current study to be superior than those not included here.


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Reviews

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Discussion

Discussion

5 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

5 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

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Hot start PCR - Mammalian DNA using KAPA LongRange from Sigma-Aldrich.

Paper title
Long-range PCR in next-generation sequencing: comparison of six enzymes and evaluation on the MiSeq sequencer
Full paper
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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for KAPA LongRange below.

Download PDF Download manufacturer protocol

Videos

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