A novel process for mutation detection using uracil DNA-glycosylase

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dc.contributor.authorVaughan, Patrick
dc.contributor.authorMcCarthy, Tommie V.
dc.contributor.funderBioResearch Ireland
dc.date.accessioned2017-11-14T13:24:33Z
dc.date.available2017-11-14T13:24:33Z
dc.date.issued1998
dc.description.abstractA novel process is presented for the detection of known mutations-and polymorphisms in DNA. This process, termed glycosylase mediated polymorphism detection (GMPD) involves amplification of the target DNA using three normal dNTPs and a fourth modified dNTP, whose base is a substrate for a specific DNA-glycosylase once incorporated into the DNA. The work described here utilises uracil DNA-glycosylase as the specific glycosylase and dUTP as the modified dNTP, Primers are designed so that during extension, the position of the first uracil incorporated into the extended primers differs depending on whether a mutation is present or absent. Subsequent glycosylase excision of the uracil residues followed by cleavage of the apyrimidinic sites allows detection of the mutation in the amplified fragment as a fragment length polymorphism. Variation in the sizes of the fragment length polymorphisms generated, can be readily achieved through the use of inosine bases in place of adenine bases in the upper and/or lower primers. The GMPD process is also adaptable to solid phase analysis. The use of the process for detection of mutations in the RYR1 and CFTR genes is demonstrated. Overall, the simplicity, specificity, versatility and flexibility of the GMPD process make it an attractive candidate for both small and large scale application in mutation detection and genome analysis.en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationVaughan, P. and McCarthy, T. V. (1998) 'A novel process for mutation detection using uracil DNA-glycosylase', Nucleic Acids Research, 26(3), pp. 810-815. doi: 10.1093/nar/26.3.810en
dc.identifier.doi10.1093/nar/26.3.810
dc.identifier.endpage815
dc.identifier.issn0305-1048
dc.identifier.issued3
dc.identifier.journaltitleNucleic Acids Researchen
dc.identifier.startpage810
dc.identifier.urihttps://hdl.handle.net/10468/5037
dc.identifier.volume26
dc.language.isoenen
dc.publisherOxford University Pressen
dc.relation.urihttps://academic.oup.com/nar/article-lookup/doi/10.1093/nar/26.3.810
dc.rights© 1998, Oxford University Pressen
dc.subjectComprehensive genetic mapen
dc.subjectCystic Fibrosis geneen
dc.subjectMalignant hyperthermiaen
dc.subjectSodium bisulfiteen
dc.subjectIdentificationen
dc.subjectAmplificationen
dc.subjectConformationen
dc.subjectCytosineen
dc.subjectGenomeen
dc.subjectAciden
dc.titleA novel process for mutation detection using uracil DNA-glycosylaseen
dc.typeArticle (peer-reviewed)en
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