{"@context":{"@vocab":"https://cir.nii.ac.jp/schema/1.0/","rdfs":"http://www.w3.org/2000/01/rdf-schema#","dc":"http://purl.org/dc/elements/1.1/","dcterms":"http://purl.org/dc/terms/","foaf":"http://xmlns.com/foaf/0.1/","prism":"http://prismstandard.org/namespaces/basic/2.0/","cinii":"http://ci.nii.ac.jp/ns/1.0/","datacite":"https://schema.datacite.org/meta/kernel-4/","ndl":"http://ndl.go.jp/dcndl/terms/","jpcoar":"https://github.com/JPCOAR/schema/blob/master/2.0/"},"@id":"https://cir.nii.ac.jp/crid/1363670319692488576.json","@type":"Article","productIdentifier":[{"identifier":{"@type":"DOI","@value":"10.1101/gr.1.1.17"}},{"identifier":{"@type":"URI","@value":"https://syndication.highwire.org/content/doi/10.1101/gr.1.1.17"}}],"dc:title":[{"@value":"DNA polymerase fidelity and the polymerase chain reaction."}],"description":[{"type":"abstract","notation":[{"@value":"<jats:p>High-fidelity DNA synthesis conditions are those that exploit the inherent ability of polymerases to discriminate against errors. This review has described several experimental approaches for controlling the fidelity of enzymatic DNA amplification. One of the most important parameters to consider is the choice of which polymerase to use in PCR. As demonstrated by the data in Tables 2 and 3, high-fidelity DNA amplification will be best achieved by using a polymerase with an active 3'-->5' proofreading exonuclease activity (Fig. 1E). For those enzymes that are proofreading-deficient, the in vitro reaction conditions can significantly influence the polymerase error rates. To maximize fidelity at the dNTP insertion step (Fig. 1A,B), any type of deoxynucleoside triphosphate pool imbalance should be avoided. Similarly, stabilization of errors by polymerase extension from mispaired or misaligned primer-termini (Fig. 1D) can be minimized by reactions using short synthesis times, low dNTP concentrations, and low enzyme concentrations. Additional improvements in fidelity can be made by further manipulating the reaction conditions. To perform high-fidelity PCR with Taq polymerase, reactions should contain a low MgCl2 concentration, not in large excess over the total concentration of dNTP substrates, and be buffered to approximately pH 6 (70 degrees C) using Bis-Tris Propane or PIPES (Table 2). These buffers have a pKa between pH 6 and pH 7 and a small temperature coefficient (delta pKa/degree C), allowing the pH to be maintained stably throughout the PCR cycle. For amplifications in which fidelity is the critical issue, one should avoid the concept that conditions generating more DNA product are the better conditions.(ABSTRACT TRUNCATED AT 250 WORDS)</jats:p>"}]}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1380004236857142548","@type":"Researcher","foaf:name":[{"@value":"K A Eckert"}]},{"@id":"https://cir.nii.ac.jp/crid/1383670319692488577","@type":"Researcher","foaf:name":[{"@value":"T A Kunkel"}]}],"publication":{"publicationIdentifier":[{"@type":"PISSN","@value":"10889051"},{"@type":"PISSN","@value":"http://id.crossref.org/issn/10889051"}],"prism:publicationName":[{"@value":"Genome Research"}],"dc:publisher":[{"@value":"Cold Spring Harbor Laboratory"}],"prism:publicationDate":"1991-08","prism:volume":"1","prism:number":"1","prism:startingPage":"17","prism:endingPage":"24"},"reviewed":"false","url":[{"@id":"https://syndication.highwire.org/content/doi/10.1101/gr.1.1.17"}],"createdAt":"2007-06-05","modifiedAt":"2024-02-14","relatedProduct":[{"@id":"https://cir.nii.ac.jp/crid/1360002220651711360","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@value":"Mutant Taq DNA polymerases with improved elongation ability as a useful reagent for genetic engineering"}]},{"@id":"https://cir.nii.ac.jp/crid/1360567183075619584","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@value":"Reduction of Werner Syndrome Protein Enhances G:C → A:T Transition by<i>O</i><sup>6</sup>-Methylguanine in Human Cells"}]},{"@id":"https://cir.nii.ac.jp/crid/1360567184619914368","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@value":"Mutations induced by 8-oxo-7,8-dihydroguanine in WRN- and DNA polymerase λ-double knockdown cells"}]},{"@id":"https://cir.nii.ac.jp/crid/1390001205474559488","@type":"Article","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@language":"en","@value":"A useful strategy to construct DNA polymerases with different properties by using genetic resources from environmental DNA"}]},{"@id":"https://cir.nii.ac.jp/crid/2050588892089660288","@type":"Article","resourceType":"学術雑誌論文(journal article)","relationType":["isReferencedBy"],"jpcoar:relatedTitle":[{"@value":"Analysis of large deletion mutations induced by abasic site analog in human cells"}]}],"dataSourceIdentifier":[{"@type":"CROSSREF","@value":"10.1101/gr.1.1.17"},{"@type":"CROSSREF","@value":"10.3389/fmicb.2014.00461_references_DOI_FqQW9XiAixwsXytBthceRWF0gNY"},{"@type":"CROSSREF","@value":"10.1186/s41021-018-0110-7_references_DOI_FqQW9XiAixwsXytBthceRWF0gNY"},{"@type":"CROSSREF","@value":"10.1266/ggs.84.3_references_DOI_FqQW9XiAixwsXytBthceRWF0gNY"},{"@type":"CROSSREF","@value":"10.1093/mutage/gey024_references_DOI_FqQW9XiAixwsXytBthceRWF0gNY"},{"@type":"CROSSREF","@value":"10.1021/acs.chemrestox.8b00009_references_DOI_FqQW9XiAixwsXytBthceRWF0gNY"}]}