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Titolo:
Quantitative determination of gene expression in human lymphocytes assessed by reverse transcription-polymerase chain reaction coupled to high-performance liquid chromatography
Autore:
Andia, I; Zumarraga, M; Davila, R; Miller, JC; Friefhoff, AJ;
Indirizzi:
Hosp Psiquiat Zamudio, Dept Invest Neuroquim, Zamudio 48170, Vizcaya, Spain Hosp Psiquiat Zamudio Zamudio Vizcaya Spain 48170 o 48170, Vizcaya, Spain NYU, Sch Med, Dept Psychiat, Millhauser Labs, New York, NY USA NYU New York NY USA ed, Dept Psychiat, Millhauser Labs, New York, NY USA
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 2, volume: 761, anno: 2001,
pagine: 237 - 246
SICI:
1387-2273(20010925)761:2<237:QDOGEI>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
NONPOROUS POLY(STYRENE-DIVINYLBENZENE) PARTICLES; COMPETITIVE RT-PCR; MESSENGER-RNA; HOMOVANILLIC-ACID; DNA FRAGMENTS; HPLC; QUANTIFICATION; ACCURATE; PLASMA;
Keywords:
gene expression; reverse transcription; polymerase chain reaction; mRNA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
28
Recensione:
Indirizzi per estratti:
Indirizzo: Andia, I Hosp Psiquiat Zamudio, Dept Invest Neuroquim, B Arteaga 107, Zamudio 48170, Vizcaya, Spain Hosp Psiquiat Zamudio B Arteaga 107 Zamudio Vizcaya Spain 48170 n
Citazione:
I. Andia et al., "Quantitative determination of gene expression in human lymphocytes assessed by reverse transcription-polymerase chain reaction coupled to high-performance liquid chromatography", J CHROMAT B, 761(2), 2001, pp. 237-246

Abstract

Gene expression in human lymphocytes was assessed using reverse transcription and polymerase chain reaction amplification followed by ion-pair reversed-phase chromatography analysis. Competitive PCR was used to quantitate the desired cDNAs with a polivalent competitor adaptable to multiple novel mRNAs estimations with minor changes. Accuracy was 11.27 +/- 11.87% (n=7), asdetermined using standards. The coefficients of variation of the assessment of human OK12b were 7% (n=6), 7.68 attmol/mug of total RNA, and 21% (n=6), 0.93 attmol/mug of total RNA. Sample-to-sample variation in the reverse transcription and in the quantity and quality of RNA was attenuated by normalising results to beta-actin mRNA expression, The correlation between the OK12b/beta -actin ratio and competitive assessments of OK12b was 0.984, n=6. The correlation between HPLC results and an independent method based on radionuclide uptake by the product, detected by electrophoretic separation, was 0.848, n=10. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/01/20 alle ore 20:29:44