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Article: Direct cycle sequencing of mutated alleles detected by PCR single-strand conformation polymorphism analysis

TitleDirect cycle sequencing of mutated alleles detected by PCR single-strand conformation polymorphism analysis
Authors
Issue Date1993
PublisherEaton Publishing Co. The Journal's web site is located at http://www.biotechniques.com
Citation
Biotechniques, 1993, v. 14 n. 5, p. 790+792-794 How to Cite?
AbstractA rapid protocol for direct sequencing of a mutated allele, detected by combining polymerase chain reaction with single-strand conformation polymorphism (PCR-SSCP) analysis and cycle sequencing using a thermal cycler, is described. End-labeled radioactive primers were used both for PCR-SSCP analysis for the detection of p53 gene mutation and for cycle sequencing using ΔTaq(TM) Version 2.0 DNA polymerase. The point mutations along the various exons of the p53 gene can be rapidly determined by this sequencing method. This protocol requires only a small amount of DNA template (0.1 μg) and produces sequencing images with low backgrounds and very uniform band intensity. It has also been used successfully in the study of other gene mutations including ras and NF-1 (neurofibromatosis 1) genes.
Persistent Identifierhttp://hdl.handle.net/10722/149532
ISSN
2015 Impact Factor: 2.298
2015 SCImago Journal Rankings: 1.132
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorMok, SCHen_US
dc.contributor.authorLo, KWen_US
dc.contributor.authorTsao, SWen_US
dc.date.accessioned2012-06-26T05:54:53Z-
dc.date.available2012-06-26T05:54:53Z-
dc.date.issued1993en_US
dc.identifier.citationBiotechniques, 1993, v. 14 n. 5, p. 790+792-794en_US
dc.identifier.issn0736-6205en_US
dc.identifier.urihttp://hdl.handle.net/10722/149532-
dc.description.abstractA rapid protocol for direct sequencing of a mutated allele, detected by combining polymerase chain reaction with single-strand conformation polymorphism (PCR-SSCP) analysis and cycle sequencing using a thermal cycler, is described. End-labeled radioactive primers were used both for PCR-SSCP analysis for the detection of p53 gene mutation and for cycle sequencing using ΔTaq(TM) Version 2.0 DNA polymerase. The point mutations along the various exons of the p53 gene can be rapidly determined by this sequencing method. This protocol requires only a small amount of DNA template (0.1 μg) and produces sequencing images with low backgrounds and very uniform band intensity. It has also been used successfully in the study of other gene mutations including ras and NF-1 (neurofibromatosis 1) genes.en_US
dc.languageengen_US
dc.publisherEaton Publishing Co. The Journal's web site is located at http://www.biotechniques.comen_US
dc.relation.ispartofBioTechniquesen_US
dc.subject.meshAllelesen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshBiotechnologyen_US
dc.subject.meshDna, Neoplasm - Chemistry - Geneticsen_US
dc.subject.meshDna, Single-Stranded - Chemistry - Geneticsen_US
dc.subject.meshFemaleen_US
dc.subject.meshGenes, P53en_US
dc.subject.meshHumansen_US
dc.subject.meshNucleic Acid Conformationen_US
dc.subject.meshOvarian Neoplasms - Geneticsen_US
dc.subject.meshPoint Mutationen_US
dc.subject.meshPolymerase Chain Reaction - Methodsen_US
dc.subject.meshPolymorphism, Geneticen_US
dc.subject.meshSequence Analysis, Dnaen_US
dc.titleDirect cycle sequencing of mutated alleles detected by PCR single-strand conformation polymorphism analysisen_US
dc.typeArticleen_US
dc.identifier.emailTsao, SW:gswtsao@hkucc.hku.hken_US
dc.identifier.authorityTsao, SW=rp00399en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid8512704-
dc.identifier.scopuseid_2-s2.0-0027252974en_US
dc.identifier.volume14en_US
dc.identifier.issue5en_US
dc.identifier.spage790+792en_US
dc.identifier.epage794en_US
dc.identifier.isiWOS:A1993LA81200029-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridMok, SCH=7006015487en_US
dc.identifier.scopusauthoridLo, KW=7402101603en_US
dc.identifier.scopusauthoridTsao, SW=7102813116en_US

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