A novel process for mutation detection using uracil DNA-glycosylase

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dc.contributor.author Vaughan, Patrick
dc.contributor.author McCarthy, Tommie V.
dc.date.accessioned 2017-11-14T13:24:33Z
dc.date.available 2017-11-14T13:24:33Z
dc.date.issued 1998
dc.identifier.citation Vaughan, 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/26.3.810 en
dc.identifier.volume 26
dc.identifier.issued 3
dc.identifier.startpage 810
dc.identifier.endpage 815
dc.identifier.issn 0305-1048
dc.identifier.uri http://hdl.handle.net/10468/5037
dc.identifier.doi 10.1093/nar/26.3.810
dc.description.abstract A 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.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Oxford University Press en
dc.relation.uri https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/26.3.810
dc.rights © 1998, Oxford University Press en
dc.subject Comprehensive genetic map en
dc.subject Cystic Fibrosis gene en
dc.subject Malignant hyperthermia en
dc.subject Sodium bisulfite en
dc.subject Identification en
dc.subject Amplification en
dc.subject Conformation en
dc.subject Cytosine en
dc.subject Genome en
dc.subject Acid en
dc.title A novel process for mutation detection using uracil DNA-glycosylase en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Tommie McCarthy, Biochemistry & Cell Biology, University College Cork, Cork, Ireland. +353-21-490-3000 Email: t.mccarthy@ucc.ie en
dc.internal.availability Full text available en
dc.description.version Published Version en
dc.contributor.funder BioResearch Ireland
dc.description.status Peer reviewed en
dc.identifier.journaltitle Nucleic Acids Research en
dc.internal.IRISemailaddress t.mccarthy@ucc.ie en


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