The fidelity of a polymerase refers to its ability to insert the correct base during PCR. Conversely, the rate of misincorporation is known as a polymerase’s error rate. High-fidelity PCR, utilizes a DNA polymerase with a low error rate and results in a high degree of accuracy in the replication of the DNA of interest. NEB scientists were the first to identify and commercialize a high-fidelity DNA polymerase suitable for PCR, namely Vent® DNA Polymerase. Since its discovery, several other high fidelity polymerases have been discovered and engineered – all with differing processivity, speed and levels of accuracy.
- PCR Protocol Phusion® DNA Polymerase
- Protocol for a Routine Deep Vent® PCR
- Protocol for Phusion® Hot Start Flex 2X Master Mix
- PCR Using NEBNext® High-Fidelity 2X PCR Master Mix (M0541)
- PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)
- Protocol for Q5® Hot Start High-Fidelity 2X Master Mix
- Guidelines for PCR Optimization for Deep Vent® DNA Polymerase
- Protocol for a Routine Deep Vent® (exo-) PCR
- PCR Protocol for Phusion® Hot Start Flex DNA Polymerase (M0535)
- Protocol Phusion® High-Fidelity PCR Master Mix with HF Buffer
- PCR Optimization with Phusion® High-Fidelity PCR Kit
- Protocol for a Routine PCR with Phusion® High-Fidelity PCR Kit
- PCR Optimization of the Control Template using Phusion® High-Fidelity PCR Kit
- Protocol for Phusion® High-Fidelity PCR Master Mix with GC Buffer
- PCR Protocol for Crimson LongAmp™ Taq DNA Polymerase (M0326)
- Protocol for Q5® High-Fidelity 2X Master Mix
- PCR Optimization (E0555)
- Protocol for a Routine PCR (E0555)
- PCR Using Q5® High-Fidelity DNA Polymerase (M0491)
Understanding Variability in DNA Amplification Reactions
Anatomy of a Polymerase - How Function and Structure are Related
Read about the relationship between Polymerase structure and function when copying DNA.
Polymerase Fidelity: What is it, and what does it mean for your PCR?
- PCR Selector
- DNA Polymerase Selection Chart
- PCR Troubleshooting Guide
- Guidelines for PCR Optimization with Thermophilic DNA Polymerases
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The secondary structures that form within GC-rich regions can make efficient DNA amplification for PCR difficult. Nicole’s suggestions can help!
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Learn how proofreading polymerases recognize and correct mismatched bases.
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Here are some quick tips for getting the most out of NEB's Q5® High-Fidelity DNA Polymerase.
Not sure why Q5® is your best choice for high-fidelity amplification of GC-rich targets? NEB's scientists will show you why we call Q5 an "ultra-high fidelity polymerase".