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!
Make sure you're using the optimal polymerase for your DNA amplifications. Get tips on choosing the right DNA Polymerase for your application.
Learn how proofreading polymerases recognize and correct mismatched bases.
Looking for tips on dealing with GC-bias in DNA amplification? NEB scientists have the expertise you need!
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".