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Titolo:
Fluorescent multiplex polymerase chain reaction analysis of four genes associated with inpaired fibrinolysis and myocardial infarction
Autore:
Renshaw, MA; Ellingsen, D; Costner, B; Benson, J; Heit, JA; Hooper, WC;
Indirizzi:
CDCP, Hematol Dis Branch, Div AIDS STD & TB Lab Res, Natl Ctr Infect Dis, Atlanta, GA 30333 USA CDCP Atlanta GA USA 30333 Res, Natl Ctr Infect Dis, Atlanta, GA 30333 USA
Titolo Testata:
BLOOD COAGULATION & FIBRINOLYSIS
fascicolo: 4, volume: 12, anno: 2001,
pagine: 245 - 251
SICI:
0957-5235(200106)12:4<245:FMPCRA>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
DNA-POLYMERASE; PCR; POLYMORPHISM; DISEASE; RISK;
Keywords:
multiplex polymerase chain reaction; angiotensin converting enzyme; tissue plasminogen activator; plasminogen activator inhibitor-1; endothelial cell nitric oxide synthase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
14
Recensione:
Indirizzi per estratti:
Indirizzo: Hooper, WC CDCP, Hematol Dis Branch, Div AIDS STD & TB Lab Res, Natl Ctr Infect Dis, MS DO2,1600 Clifton Rd, Atlanta, GA 30333 USA CDCP MS DO2,1600 Clifton Rd Atlanta GA USA 30333 , GA 30333 USA
Citazione:
M.A. Renshaw et al., "Fluorescent multiplex polymerase chain reaction analysis of four genes associated with inpaired fibrinolysis and myocardial infarction", BL COAG FIB, 12(4), 2001, pp. 245-251

Abstract

Mltiplex polymerase chain reaction (PCR) allows for the simultaneous amplification of several genes, thereby optimizing the use of reagents and decreasing personnel time. Multiplex PCR was used to amplify four genes in one PCR reaction, demonstrating the advantage of multiplex PCR for our study since it allowed us to amplify four separate genes using only 1 mul DNA, thus maximizing the use of study DNA. As compared with conventional multiplex PCR analysis with ethidium bromide, incorporating fluorescence-labeled primers into multiplex PCR reactions facilitated accurate, simultaneous analysis of many DNA fragments within one base discrimination. We have used this fluorescence methodology to analyze polymorphisms associated with either impaired fibrinolysis or myocardial infarction. These include the angiotensin converting enzyme insertion/deletion (I/D) polymorphism in intron 16 of the DCP1 gene, the Alu I/D polymorphism of the tissue plasminogen activator-25 locus in intron 8, the plasminogen activator inhibitor 4G/5G repeat polymorphism, and the variable number tandem repeat of the endothelial cell nitric oxide synthase gene, all characterized by an insertion, deletion, or repeat. The amplified products were diluted 1 :60 and analyzed on the ABI PRISMN 310 Genetic Analyzer using GeneScan(N) software. With this method, we were able to amplify four genes using 75% less reagents and personnel time, thusdemonstrating the benefit of multiplex PCR and fluorescence technology. (C) 2001 Lippincott Williams & ilkins.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 11/07/20 alle ore 21:11:32