Source:http://linkedlifedata.com/resource/pubmed/id/17105184
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
22
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pubmed:dateCreated |
2006-11-19
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pubmed:abstractText |
DNA diagnostic has been moving from expensive, low-throughput, multistep methods to inexpensive, higher throughput, closed-tube, and automated methods. Fluorescence is the favored signaling technology for such assays. In this method, we describe a universal molecular beacon (U-MB) as the fluorescent tracer in the real-time PCR technique. A 5'-universal template primer (5'-UT primer) has been designed with a tail in complementary to the loop and 5'-side arm sequence of U-MB at the 5'-end of forward target specific primer. As PCR cycles increase, a new DNA fragment with a 5'-UT primer tail is synthesized, which is used as the template for next PCR cycle. As the reverse primer extends to the 5'-UT primer tail, the U-MB hybridized is displaced and the fluorescence from the fluorophore of the U-MB is quenched, indicating that the allele-specific PCR is in progress. This tracing system combined with an allele-specific reverse primer and vent (exo-) DNA polymerase, a polymerase that lacks 3'- to 5'-exonuclease activity, was used for the detection of point mutations of base G in codon 259 (AGA) of exon 7 of p53 gene on a panel of breast cancer individuals.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Codon,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Directed RNA Polymerases,
http://linkedlifedata.com/resource/pubmed/chemical/Oligonucleotide Probes,
http://linkedlifedata.com/resource/pubmed/chemical/RNA,
http://linkedlifedata.com/resource/pubmed/chemical/RNA primers,
http://linkedlifedata.com/resource/pubmed/chemical/Tumor Suppressor Protein p53
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0003-2700
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
78
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
7886-90
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:17105184-Breast Neoplasms,
pubmed-meshheading:17105184-Codon,
pubmed-meshheading:17105184-DNA Primers,
pubmed-meshheading:17105184-DNA-Directed RNA Polymerases,
pubmed-meshheading:17105184-Exons,
pubmed-meshheading:17105184-Female,
pubmed-meshheading:17105184-Genetic Testing,
pubmed-meshheading:17105184-Genotype,
pubmed-meshheading:17105184-Humans,
pubmed-meshheading:17105184-Mutation,
pubmed-meshheading:17105184-Oligonucleotide Probes,
pubmed-meshheading:17105184-Polymerase Chain Reaction,
pubmed-meshheading:17105184-RNA,
pubmed-meshheading:17105184-Templates, Genetic,
pubmed-meshheading:17105184-Tumor Suppressor Protein p53
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pubmed:year |
2006
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pubmed:articleTitle |
Universal molecular beacon-based tracer system for real-time polymerase chain reaction.
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pubmed:affiliation |
The Key Laboratory of Bioorganic Chemistry & Molecular Engineering and Institute of Analytical Chemistry, College of Chemistry & Molecular Engineering, Peking University, Beijing, 100871, China.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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