Linked Life Data

11513552

Source:http://linkedlifedata.com/resource/pubmed/id/11513552

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2001-8-21
pubmed:abstractText
Relative quantitative reverse transcription-polymerase chain reaction (rqRT-PCR), which allows an accurate quantification of the amount of mRNA in samples potentially differing in the quality of their RNA preparation, was used to quantify lipoprotein lipase (LPL) mRNA in ovine adipose tissue. A comparative evaluation of four rqRT-PCR procedures was carried out. The amount of LPL mRNA was assayed relative to either that of gamma-actin (ACT) or cyclophilin (CYC) mRNA, used as endogenous standard. Independent (INACT and INCYC procedures) or simultaneous (COACT and COCYC procedures) amplifications have been compared. Fluorescently labelled primers yielded PCR products which were quantitatively analysed using an automated DNA sequencer. After optimizing the PCR cycle number and verifying that the amounts of ACT and CYC mRNA varied only weakly according to the nutritional conditions studied, we have tested the ability of the four procedures to quantify specific variations in LPL mRNA. The repeatability of each step and the overall assay reproducibility were also examined. The COACT and INCYC procedures were finally retained to accurately quantify LPL mRNA in AT from nine underfed or refed ewes, and gave highly correlated results (r=0.98, p<0.01). In addition, significant correlations (r=0.83, p<0.01 and r=0.92, p<0.01 for COACT and INCYC, respectively) were observed with the LPL activity in AT.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0890-8508
pubmed:author
pubmed:copyrightInfo
Copyright 2001 Academic Press.
pubmed:issnType
Print
pubmed:volume
15
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
187-94
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
2001
pubmed:articleTitle
A fluorescent reverse transcription-polymerase chain reaction assay to quantify the lipoprotein lipase messenger RNA.
pubmed:affiliation
INRA, Unité de Recherches sur les Herbivores, Saint-Genes-Champanelle, 63122, France. mbonnet@clermont.inra.fr
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't