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)
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?
Understanding Variability in DNA Amplification Reactions
- PCR Selector
- DNA Polymerase Selection Chart
- PCR Troubleshooting Guide
- Guidelines for PCR Optimization with Thermophilic DNA Polymerases
- Vladimir Potapov, Jennifer L. Ong. 2017. Examining Sources of Error in PCR by Single-Molecule Sequencing. PLOS One. , PubMedID: 28683110, DOI:
- Yafeng Li, Delu Song, Ying Song, Liangliang Zhao, Natalie Wolkow, John W Tobias, Wenchao Song, Joshua L Dunaief 2015. Iron-induced Local Complement Component 3 (C3) Up-regulation via Non-canonical Transforming Growth Factor (TGF)-β Signaling in the Retinal Pigment Epithelium. J Biol Chem. 290, PubMedID: 25802332, DOI: 10.1074/jbc.M115.645903
- Yuan Xue, Jossef Osborn, Anand Panchal, Jay L Mellies 2015. The RpoE Stress Response Pathway Mediates Reduction of the Virulence of Enteropathogenic Escherichia coli by Zinc. Appl Environ Microbiol. 81, PubMedID: 25819956, DOI: 10.1128/AEM.00507-15
- Longhai Dai, Can Liu, Yueming Zhu, Jiangsheng Zhang, Yan Men, Zeng Yan, Yuanxia Sun 2015. Functional Characterization of Cucurbitadienol Synthase and Triterpene Glycosyltransferase Involved in Biosynthesis of Mogrosides from Siraitia grosvenorii. Plant Cell Physiol. , PubMedID: 25759326, DOI: 10.1093/pcp/pcv043
- Harish Nag Kankipati, Marta Rubio-Texeira, Dries Castermans, George Diallinas, Johan M Thevelein 2015. Sul1 and Sul2 Sulfate Transceptors Signal to Protein Kinase A upon Exit of Sulfur Starvation. J Biol Chem. 290, PubMedID: 25724649, DOI: 10.1074/jbc.M114.629022
- Amin Zargar, David N Quan, Milad Emamian, Chen Yu Tsao, Hsuan-Chen Wu, Chelsea R Virgile, William E Bentley 2015. Rational design of 'controller cells' to manipulate protein and phenotype expression. Metab Eng. , PubMedID: 25908186, DOI: 10.1016/j.ymben.2015.04.001
- Christine Henke, Pamela L Strissel, Maria-Theresa Schubert, Megan Mitchell, Claus C Stolt, Florian Faschingbauer, Matthias W Beckmann, Reiner Strick 2015. Selective expression of sense and antisense transcripts of the sushi-ichi-related retrotransposon - derived family during mouse placentogenesis. Retrovirology. 12, PubMedID: 25888968, DOI: 10.1186/s12977-015-0138-8
- Yonghe Zhang, Huiming Huang, Shanshan Xu, Bo Wang, Jianhua Ju, Huarong Tan, Wenli Li 2015. Activation and enhancement of Fredericamycin A production in deepsea-derived Streptomyces somaliensis SCSIO ZH66 by using ribosome engineering and response surface methodology. Microb Cell Fact. 14, PubMedID: 25927229, DOI: 10.1186/s12934-015-0244-2
- Binyamin D Berkovits, Christine Mayr 2015. Alternative 3' UTRs act as scaffolds to regulate membrane protein localization. Nature. , PubMedID: 25896326, DOI: 10.1038/nature14321
- Jun Wu, Daiji Okamura, Mo Li, Keiichiro Suzuki, Chongyuan Luo, Li Ma, Yupeng He, Zhongwei Li, Chris Benner, Isao Tamura, Marie N Krause, Joseph R Nery, Tingting Du, Zhuzhu Zhang, Tomoaki Hishida, Yuta Takahashi, Emi Aizawa, Na Young Kim, Jeronimo Lajara, Pedro Guillen, Josep M Campistol, Concepcion Rodriguez Esteban, Pablo J Ross, Alan Saghatelian, Bing Ren, Joseph R Ecker, Juan Carlos Izpisua Belmonte 2015. An alternative pluripotent state confers interspecies chimaeric competency. Nature. , PubMedID: 25945737, DOI: 10.1038/nature14413
- Bert De Rybel, Milad Adibi, Alice S. Breda, Jos R. Wendrich, Margot E. Smit, Ondej Novk, Nobutoshi Yamaguchi, Saiko Yoshida, Gert Van Isterdael, Joakim Palovaara, Bart Nijsse, Mark V. Boekschoten, Guido Hooiveld, Tom Beeckman, Doris Wagner, Karin Ljung, Christian Fleck, Dolf Weijers 2014. Integration of growth and patterning during vascular tissue formation in Arabidopsis Science. 345, PubMedID: 25104393, DOI: 10.1126/science.1255215
- Xin Duan, Arjun Krishnaswamy, Irina De la Huerta, Joshua R Sanes 2014. Type II Cadherins Guide Assembly of a Direction-Selective Retinal Circuit. Cell. 158, PubMedID: 25126785, DOI: 10.1016/j.cell.2014.06.047
- Martin Kostovcik, Craig C Bateman, Miroslav Kolarik, Lukasz L Stelinski, Bjarte H Jordal, Jiri Hulcr 2014. The ambrosia symbiosis is specific in some species and promiscuous in others: evidence from community pyrosequencing. ISME J. , PubMedID: 25083930, DOI: 10.1038/ismej.2014.115
- Vidhyadhar Nandana, Sushant Singh, Abhay Narayan Singh, Vikash Kumar Dubey 2014. Procerain B, a cysteine protease from Calotropis procera, requires N-terminus pro-region for activity: cDNA cloning and expression with pro-sequence. Protein Expr Purif. 103C, PubMedID: 25173974, DOI: 10.1016/j.pep.2014.08.003
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
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".