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Titolo:
Quantitative TaqMan PCR without a real-time thermal cycler: An assay for fish insulin-like growth factor I messenger RNA
Autore:
Dyer, A; Soole, K; Matsumoto, G;
Indirizzi:
Cooperat Ctr Tissue Growth & Repair, Bedford Pk, SA 5042, Australia Cooperat Ctr Tissue Growth & Repair Bedford Pk SA Australia 5042 ustralia Flinders Univ S Australia, Sch Biol Sci, Bedford Pk, SA 5042, Australia Flinders Univ S Australia Bedford Pk SA Australia 5042 SA 5042, Australia Monterey Bay Aquarium Res Inst, Moss Landing, CA 95039 USA Monterey Bay Aquarium Res Inst Moss Landing CA USA 95039 ng, CA 95039 USA
Titolo Testata:
MARINE BIOTECHNOLOGY
fascicolo: 1, volume: 3, anno: 2001,
pagine: 16 - 21
SICI:
1436-2228(200101/02)3:1<16:QTPWAR>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE-CHAIN-REACTION; RT-PCR; QUANTIFICATION; EXPRESSION;
Keywords:
insulin-like growth factor I; messenger RNA; RT-PCR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
13
Recensione:
Indirizzi per estratti:
Indirizzo: Dyer, A Cooperat Ctr Tissue Growth & Repair, Sturt Rd, Bedford Pk, SA 5042, Australia Cooperat Ctr Tissue Growth & Repair Sturt Rd Bedford Pk SA Australia 5042
Citazione:
A. Dyer et al., "Quantitative TaqMan PCR without a real-time thermal cycler: An assay for fish insulin-like growth factor I messenger RNA", MAR BIOTEC, 3(1), 2001, pp. 16-21

Abstract

The reverse transcriptase-polymerase chain reaction, more commonly known as RT-PCR, has become a widely used tool in molecular biology and is now frequently used in monitoring gene expression levels. A number of variations in the RT-PCR technique now exist including TaqMan PCR (5' nuclease assay), which is a useful nonisotopic detection method for the quantification of PCR products. To monitor the formation of these fluorescent amplification products a "real-time" thermal cycler is normally required. In this study, repeated scanning of PCR products in a 96-well plate format showed that a conventional fluorescent plate reader can be used to generate similar results. To demonstrate the power of this approach, the nutritional regulation of insulin-like growth factor I (IGF-I) was investigated in a marine finfish, the snapper (Pagrus auratus). Hepatic IGF-I messenger RNA levels were shown to significantly decrease after 2 weeks of fasting and returned to fed control levels on refeeding. These results demonstrated that a real-time PCR machine was not required to generate this type of quantitative data and that this technology can be adapted for use in most molecular biology laboratories.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 22/01/20 alle ore 09:40:00