|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
8
|
|
pubmed:dateCreated |
2000-4-10
|
|
pubmed:abstractText |
A novel signal generation principle suitable for real time and end-point detection of specific PCR products in a closed tube is described. Linear DNA probes were labeled at their 5'-ends with a stable, fluorescent terbium chelate. The fluorescence intensity of this chelate is lower when it is coupled to single-stranded DNA than when the chelate is free in solution. The synthesized probes were used in the real time monitoring of PCR using a prototype instrument that consisted of a fluorometer coupled to a thermal cycler. When the probe anneals to a complementary target amplicon, the 5'-->3' exonucleolytic activity of DNA polymerase detaches the label from the probe. This results in an enhanced terbium fluorescence signal. Since terbium has a long excited state lifetime, its fluorescence can be measured in a time-resolved manner, which results in a low background fluorescence and a 1000-fold signal amplification. The detection method is quantitative over an extremely wide linear range (at least 10-10(7)initial template molecules). The label strategy can easily be combined with existing label technologies, such as TaqMan 5'-exonuclease assays, in order to carry out multiplex assays that do not suffer from overlapping emission peaks of the fluorophores.
|
|
pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10047506,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10102903,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10339598,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10397261,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10429248,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10471641,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-10545062,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-1368485,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-1406315,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-1871133,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-2436946,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-2999980,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-7580930,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-7752929,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-8041624,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-8367293,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-8994660,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-8994665,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-9171107,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-9447593,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-9630890,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-9676381,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10734205-9894600
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Apr
|
|
pubmed:issn |
1362-4962
|
|
pubmed:author |
|
|
pubmed:issnType |
Electronic
|
|
pubmed:day |
15
|
|
pubmed:volume |
28
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
E28
|
|
pubmed:dateRevised |
2009-11-18
|
|
pubmed:meshHeading |
pubmed-meshheading:10734205-Chelating Agents,
pubmed-meshheading:10734205-DNA, Complementary,
pubmed-meshheading:10734205-DNA Probes,
pubmed-meshheading:10734205-DNA-Directed DNA Polymerase,
pubmed-meshheading:10734205-Feasibility Studies,
pubmed-meshheading:10734205-Fluorescent Dyes,
pubmed-meshheading:10734205-Fluorometry,
pubmed-meshheading:10734205-Humans,
pubmed-meshheading:10734205-Mutation,
pubmed-meshheading:10734205-Nucleic Acids,
pubmed-meshheading:10734205-Plasmids,
pubmed-meshheading:10734205-Polymerase Chain Reaction,
pubmed-meshheading:10734205-Prostate-Specific Antigen,
pubmed-meshheading:10734205-Terbium
|
|
pubmed:year |
2000
|
|
pubmed:articleTitle |
A new label technology for the detection of specific polymerase chain reaction products in a closed tube.
|
|
pubmed:affiliation |
Department of Biotechnology, Tykistökatu 6A, 6th Floor, University of Turku, 20520 Turku, Finland. jussi.nurmi@utu.fi
|
|
pubmed:publicationType |
Journal Article
|
