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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1997-9-25
|
| pubmed:abstractText |
The first application of frequency-domain, on-the-fly fluorescence lifetime detection to the detection, identification and resolution of fluorescent labeled oligonucleotide primers in capillary electrophoresis is presented. These studies, which are directed towards the development of four-decay DNA sequencing, employed two common sequencing primers, SP6 and M13/pUC. The primers were each tagged by a derivative of either fluorescein or BODIPY. Fluorescence emission maxima and lifetimes of the free dyes and the labeled primers were determined both in batch mode and on-the-fly in capillary electrophoresis. Fluorescence intensity and lifetime electropherograms were extracted from dynamic lifetime data that were acquired at 0.1-s intervals during the CE separation. Lifetimes were recovered using either conventional non-linear least-squares analysis or the self-modeling maximum entropy method, which does not require prior knowledge of the system. Based on migration time and fluorescence lifetime, peaks could be detected and identified, and co-eluting peaks could be resolved. Interference from background impurities and scattered light was greatly reduced by the combination of physical separation and lifetime resolution.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jul
|
| pubmed:issn |
1387-2273
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
18
|
| pubmed:volume |
695
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
85-92
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading | |
| pubmed:year |
1997
|
| pubmed:articleTitle |
On-the-fly fluorescence lifetime detection of labeled DNA primers.
|
| pubmed:affiliation |
Department of Chemistry, P.M. Gross Chemical Laboratory, Duke University, Durham, NC 27708-0346, USA.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|