| DHPLC in clinical molecular diagnostic services. | |
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MedLine Citation:
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PMID: 16202954 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A high-capacity low-cost mutation scanning method based on denaturing high-performance liquid chromatography (DHPLC) has been recently introduced. We have implemented an automated and cost-effective strategy using DHPLC. To facilitate the semi-automated analysis of multiple exons, two steps were taken. The first step was the development of a PCR protocol for the amplification of multiple exons under the same conditions. Primer sets, which amplify each exon in the entire gene, were aliquoted to and air-dried on a 96-well format PCR plate. In this way, all the exons in a gene can be simultaneously amplified on a single PCR machine. The second step was the serial DHPLC analysis of multiple amplicons under conditions optimal for each amplicon. We named the 96-well plate containing the primer pairs and the corresponding computer file used to analyze each amplicon under the pre-determined optimal conditions as the "Condition-Oriented-PCR primer-Embedded-Reactor plate," or the COPPER plate. We have developed COPPER plate systems for more than 20 congenital disorders including classic congenital syndromes like Marfan syndrome (FBN1: 65 amplicons), CHARGE syndrome (CHD7: 39 amplicons), de Lange syndrome (NIPBL: 46 amplicons), Sotos syndrome (NSD1: 30 amplicons), and Rubinstein-Taybi syndrome (CREBBP: 41 amplicons). Using the COPPER plate system, we are functioning as a reference laboratory for the clinical molecular diagnosis of congenital malformation syndromes and are presently analyzing more than 200 samples annually from all over Japan. |
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Authors:
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Kenjiro Kosaki; Toru Udaka; Torayuki Okuyama |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Review |
Journal Detail:
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Title: Molecular genetics and metabolism Volume: 86 ISSN: 1096-7192 ISO Abbreviation: Mol. Genet. Metab. Publication Date: 2005 Sep-Oct |
Date Detail:
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Created Date: 2005-10-05 Completed Date: 2006-01-23 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9805456 Medline TA: Mol Genet Metab Country: United States |
Other Details:
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Languages: eng Pagination: 117-23 Citation Subset: IM |
Affiliation:
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Department of Pediatrics, Keio University School of Medicine, Tokyo, Japan. kkosaki@sc.itc.keio.ac.jp |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Automation Chromatography, High Pressure Liquid / methods* Nucleic Acid Denaturation Polymerase Chain Reaction / instrumentation |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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