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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
21
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pubmed:dateCreated |
2004-12-3
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pubmed:abstractText |
Here we present MethylQuant, a novel method that allows accurate quantification of the methylation level of a specific cytosine within a complex genome. This method relies on the well-established treatment of genomic DNA with sodium bisulfite, which converts cytosine into uracil without modifying 5-methyl cytosine. The region of interest is then PCR-amplified and quantification of the methylation status of a specific cytosine is performed by methylation-specific real-time PCR with SYBR Green I using one of the primers whose 3' end discriminates between the methylation states of this cytosine. The presence of a locked nucleic acid at the 3' end of the discriminative primer provides the specificity necessary for accurate and sensitive quantification, even when one of the methylation states is present at a level as low as 1% of the overall population. We demonstrate that accurate quantification of the methylation status of specific cytosines can be achieved in biological samples. The method is high-throughput, cost-effective, relatively simple and does not require any specific equipment other than a real-time PCR instrument.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-10579942,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-10734209,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-10933850,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-11296230,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-11311228,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-11951649,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12042769,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12238773,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12481262,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12610534,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12671664,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12758266,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12815597,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-12866415,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-14722226,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-1542678,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-4080553,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-8790415,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-9016686,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-9171109,
http://linkedlifedata.com/resource/pubmed/commentcorrection/15576675-9336479
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/5-Methylcytosine,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Neoplasm,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescent Dyes,
http://linkedlifedata.com/resource/pubmed/chemical/Oligonucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Oligonucleotides, Antisense,
http://linkedlifedata.com/resource/pubmed/chemical/Organic Chemicals,
http://linkedlifedata.com/resource/pubmed/chemical/SYBR Green I,
http://linkedlifedata.com/resource/pubmed/chemical/Sulfites,
http://linkedlifedata.com/resource/pubmed/chemical/locked nucleic acid,
http://linkedlifedata.com/resource/pubmed/chemical/sodium bisulfite
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pubmed:status |
MEDLINE
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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:volume |
32
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
e168
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:15576675-5-Methylcytosine,
pubmed-meshheading:15576675-Animals,
pubmed-meshheading:15576675-Cell Line, Tumor,
pubmed-meshheading:15576675-CpG Islands,
pubmed-meshheading:15576675-DNA, Neoplasm,
pubmed-meshheading:15576675-DNA Methylation,
pubmed-meshheading:15576675-DNA Primers,
pubmed-meshheading:15576675-Fluorescent Dyes,
pubmed-meshheading:15576675-Genomics,
pubmed-meshheading:15576675-Oligonucleotides,
pubmed-meshheading:15576675-Oligonucleotides, Antisense,
pubmed-meshheading:15576675-Organic Chemicals,
pubmed-meshheading:15576675-Polymerase Chain Reaction,
pubmed-meshheading:15576675-Rats,
pubmed-meshheading:15576675-Sequence Analysis, DNA,
pubmed-meshheading:15576675-Sulfites
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pubmed:year |
2004
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pubmed:articleTitle |
MethylQuant: a sensitive method for quantifying methylation of specific cytosines within the genome.
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pubmed:affiliation |
Institut Jacques Monod du CNRS, Universités Paris 6-7, Tour 43, 2 place Jussieu, 75251 Paris Cedex 05, France.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Evaluation Studies
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