KAPA LongRange
PCR Hot start PCR - Mammalian DNA
Protocol tips
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| This product can also be used to amplify mid to long-range targets (up to 15 kb) |
Publication protocol
Enzymes and AmpliconsWe 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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Discussion
Discussion
5 years ago
Author:
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
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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Manufacturer protocol
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