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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
11
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pubmed:dateCreated |
2003-5-28
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pubmed:abstractText |
Here we report proof-of-principle for a microsphere-based genotyping assay that detects single nucleotide polymorphisms (SNPs) directly from human genomic DNA samples. This assay is based on a structure-specific cleavage reaction that achieves single base discrimination with a 5'-nuclease which recognizes a tripartite substrate formed upon hybridization of target DNA with probe and upstream oligonucleotides. The assay is simple with two easy steps: a cleavage reaction, which generates fluorescent signal on microsphere surfaces, followed by flow cytometry analysis of the microspheres. Genomic DNA samples were genotyped for the SNP in the Apolipoprotein E gene at amino acid position 158. The assay successfully scored wild type, heterozygous and homozygous mutants. To our knowledge, this is the first report of a solid-support assay for detection of SNPs directly from genomic DNA without PCR amplification of the target.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/12771230-10096299,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/12771230-10679731,
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
1362-4962
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:day |
1
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pubmed:volume |
31
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
e66
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:12771230-Apolipoproteins E,
pubmed-meshheading:12771230-Flow Cytometry,
pubmed-meshheading:12771230-Fluorescent Dyes,
pubmed-meshheading:12771230-Genome, Human,
pubmed-meshheading:12771230-Genotype,
pubmed-meshheading:12771230-Humans,
pubmed-meshheading:12771230-Microspheres,
pubmed-meshheading:12771230-Oligonucleotides,
pubmed-meshheading:12771230-Polymorphism, Single Nucleotide,
pubmed-meshheading:12771230-Sequence Analysis, DNA
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pubmed:year |
2003
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pubmed:articleTitle |
Genotyping single nucleotide polymorphisms directly from genomic DNA by invasive cleavage reaction on microspheres.
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pubmed:affiliation |
Department of Biomedical Engineering, Robert R. McCormick School of Engineering and Applied Science, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208-3107, USA.
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pubmed:publicationType |
Journal Article,
Evaluation Studies
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