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DHPLC analysis of adenomatous polyposis coli (APC) mutations using ready-to-use APC plates: simple detection of multiple base pair deletion mutations

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dc.contributor.authorKim, Il-Jin-
dc.contributor.authorKim, Kun-
dc.contributor.authorKang, Hio Chung-
dc.contributor.authorJang, Sang-Geun-
dc.contributor.authorPark, Jae-Gahb-
dc.date.accessioned2010-06-24T05:14:48Z-
dc.date.available2010-06-24T05:14:48Z-
dc.date.issued2008-06-17-
dc.identifier.citationGenet Test. 12(2):295-298en
dc.identifier.issn1090-6576 (Print)-
dc.identifier.urihttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18554166-
dc.identifier.urihttp://hdl.handle.net/10371/67775-
dc.description.abstractThe adenomatous polyposis coli (APC), which is the susceptible gene for familial adenomatous polyposis (FAP) and sporadic colorectal cancer, spans 15 exons. The open reading frame of APC is 8529 bp, which encodes 2843 amino acids. Conventional genetic screening involves extensive time as well as high cost and labor. Thus, we developed a novel APC ready-to-use plate for high-throughput mutational analysis by denaturing high performance liquid chromatography (DHPLC). To prepare the ready-to-use APC plate, all 38 primer pairs and PCR mixtures were aliquoted into individual wells of a 96-well plate, and frozen at -20 degrees C until use. All 38 PCR primers were designed to be amplified at the same temperature (52 degrees C). We examined a total of 27 FAP patient samples with APC germline mutations (17 for multiple bp deletions, 1 for 1 bp deletion, 9 for nonsense mutations) and 50 APC-negative noncarriers. All 17 multiple bp deletion mutations were detected during the initial 50 degrees C running analysis and thus ruled out for further analyses. All other mutations were clearly detected under specific optimized conditions. More than 50% of the APC germline mutations were multiple base pair deletions and efficiently selected by omitting time-consuming partial denaturing conditions.en
dc.language.isoen-
dc.publisherMary Ann Lieberten
dc.subjectAdenomatous Polyposis Coli/*geneticsen
dc.subjectAdenomatous Polyposis Coli Protein/*geneticsen
dc.subjectBase Pairing/*geneticsen
dc.subjectChromatography, High Pressure Liquid/*methodsen
dc.subjectDNA Mutational Analysisen
dc.subjectExonsen
dc.subject*Gene Deletionen
dc.subjectGenetic Screening/methodsen
dc.subjectHumansen
dc.subject*Mutationen
dc.titleDHPLC analysis of adenomatous polyposis coli (APC) mutations using ready-to-use APC plates: simple detection of multiple base pair deletion mutationsen
dc.typeArticleen
dc.contributor.AlternativeAuthor김건-
dc.contributor.AlternativeAuthor김일진-
dc.contributor.AlternativeAuthor강효정-
dc.contributor.AlternativeAuthor장상근-
dc.contributor.AlternativeAuthor박재갑-
dc.identifier.doi10.1089/gte.2007.0104-
dc.identifier.doi10.1089/gte.2007.0104-
Appears in Collections:
College of Medicine/School of Medicine (의과대학/대학원)Surgery (외과학전공)Journal Papers (저널논문_외과학전공)
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