| MeltMADGE for mutation scanning of specific genes in population studies. | |
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MedLine Citation:
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PMID: 21030955 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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MeltMADGE reconfigures the mutation scanning process of denaturing gradient gel electrophoresis so that the independent variable is time rather than space and the dependent (denaturing) variable is temperature rather than concentration of chemical denaturant. Use of a thermal ramp enables the use of a homogeneous gel and therefore of high-density arrays of wells such as those of microplate array diagonal gel electrophoresis (MADGE). In this configuration, electrophoresis of products on 10-12 96-well meltMADGE gels can be conducted in a 1- to 2-liter tank in a 1- to 2-h run, enabling the scanning of a target amplicon in over 1,000 subjects simultaneously. Gels are read by imaging the fluorescence of UV-excited ethidium bromide, giving a simple, economical system for identifying rarer sequence variants in target genes; it is suitable for large-scale case-control or population studies and other comparable applications. Different amplicons with similar melting characteristics can also be combined in the same run. |
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Authors:
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Khalid K Alharbi; Mohammed A Aldahmesh; Tom R Gaunt; Hamid Rassoulian; Philip Ai Guthrie; Santiago Rodriguez; Christopher R Boustred; Emmanuel Spanakis; Ian N M Day |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-10-21 |
Journal Detail:
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Title: Nature protocols Volume: 5 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2010 Nov |
Date Detail:
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Created Date: 2010-10-29 Completed Date: 2011-01-21 Revised Date: 2013-04-18 |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 1800-12 Citation Subset: IM |
Affiliation:
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Clinical Laboratory Sciences Department, College of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals DNA Mutational Analysis / instrumentation, methods* Denaturing Gradient Gel Electrophoresis Electrophoresis, Polyacrylamide Gel / instrumentation, methods* Humans Nucleic Acid Hybridization Polymerase Chain Reaction Time |
| Grant Support | |
ID/Acronym/Agency:
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086456//Wellcome Trust; //Medical Research Council; //Department of Health; //Wellcome Trust; //British Heart Foundation |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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