Pwo Polymerase Error Rate
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High Fidelity Polymerase
May 2014; Accepted 21 July 2014; Published 17 August 2014Academic Editor: Alessandro Desideri Copyright © 2014 Peter McInerney et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.AbstractAs larger-scale cloning projects become more prevalent, there is an increasing need for comparisons among high fidelity DNA polymerases used for PCR amplification. All polymerases marketed for PCR applications are tested for fidelity properties (i.e., error rate determination) by vendors, and numerous literature reports have addressed PCR enzyme fidelity. Nonetheless, it is often difficult to make direct comparisons among different enzymes due to numerous methodological and analytical differences from study to study. We have measured the error rates for 6 DNA polymerases commonly used in PCR applications, including 3 polymerases typically used for cloning applications requiring high fidelity. Error rate measurement values reported here were obtained by direct sequencing of cloned PCR products. The strategy employed here allows interrogation of error rate across a very large DNA sequence spa
Download Full-text PDF Error Rate Comparison during Polymerase Chain Reaction by DNA PolymeraseArticle (PDF Available) · August 2014 with 162 ReadsDOI: 10.1155/2014/287430 ·
Q5 Vs Phusion
Source: PubMed1st Peter McInerney2nd Paul D Adams46.23 · Lawrence Berkeley National fidelity of dna replication Laboratory3rd Masood Z HadiAbstractAs larger-scale cloning projects become more prevalent, there is an increasing need for pcr polymerase comparisons among high fidelity DNA polymerases used for PCR amplification. All polymerases marketed for PCR applications are tested for fidelity properties (i.e., error rate determination) by https://www.hindawi.com/journals/mbi/2014/287430/ vendors, and numerous literature reports have addressed PCR enzyme fidelity. Nonetheless, it is often difficult to make direct comparisons among different enzymes due to numerous methodological and analytical differences from study to study. We have measured the error rates for 6 DNA polymerases commonly used in PCR applications, including 3 polymerases typically used https://www.researchgate.net/publication/265418094_Error_Rate_Comparison_during_Polymerase_Chain_Reaction_by_DNA_Polymerase for cloning applications requiring high fidelity. Error rate measurement values reported here were obtained by direct sequencing of cloned PCR products. The strategy employed here allows interrogation of error rate across a very large DNA sequence space, since 94 unique DNA targets were used as templates for PCR cloning. The six enzymes included in the study, Taq polymerase, AccuPrime-Taq High Fidelity, KOD Hot Start, cloned Pfu polymerase, Phusion Hot Start, and Pwo polymerase, we find the lowest error rates with Pfu, Phusion, and Pwo polymerases. Error rates are comparable for these 3 enzymes and are >10x lower than the error rate observed with Taq polymerase. Mutation spectra are reported, with the 3 high fidelity enzymes displaying broadly similar types of mutations. For these enzymes, transition mutations predominate, with little bias observed for type of transition.Discover the world's research11+ million members100+ million publications100k+ research projectsJoin for free Full-text (PDF)DOI: ·Available from: PubMed Central, Dec 26, 2014 · License: C
during Polymerase Chain Reaction by DNA Polymerase. Mol Biol Int Mol Biol Int 2014 17;2014:287430. Epub 2014 Dec http://www.pubfacts.com/detail/25197572/Error-Rate-Comparison-during-Polymerase-Chain-Reaction-by-DNA-Polymerase 17. Peter McInerney, Paul Adams, Masood Z Hadi Download Full Paper As larger-scale cloning projects become more prevalent, there is an increasing need for comparisons among high fidelity DNA polymerases used for PCR amplification. All polymerases marketed for PCR applications are tested for fidelity properties (i.e., error rate determination) by vendors, and error rate numerous literature reports have addressed PCR enzyme fidelity. Nonetheless, it is often difficult to make direct comparisons among different enzymes due to numerous methodological and analytical differences from study to study. We have measured the error rates for 6 DNA polymerases commonly used in PCR applications, including 3 polymerases typically used for pwo polymerase error cloning applications requiring high fidelity. Error rate measurement values reported here were obtained by direct sequencing of cloned PCR products. The strategy employed here allows interrogation of error rate across a very large DNA sequence space, since 94 unique DNA targets were used as templates for PCR cloning. The six enzymes included in the study, Taq polymerase, AccuPrime-Taq High Fidelity, KOD Hot Start, cloned Pfu polymerase, Phusion Hot Start, and Pwo polymerase, we find the lowest error rates with Pfu, Phusion, and Pwo polymerases. Error rates are comparable for these 3 enzymes and are >10x lower than the error rate observed with Taq polymerase. Mutation spectra are reported, with the 3 high fidelity enzymes displaying broadly similar types of mutations. For these enzymes, transition mutations predominate, with little bias observed for type of transition. Affiliation Joint BioEnergy Institute, Emeryville, CA, USA ; Physical Biosciences Division, Lawrence Berkeley National Laboratories, Berkeley, CA 94720, USA ; Synthe