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Titolo:
Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-embedded tissues
Autore:
Sato, Y; Sugie, R; Tsuchiya, B; Kameya, T; Natori, M; Mukai, K;
Indirizzi:
Kitasato Univ, Sch Allied Hlth Sci, Dept Mol Diagnost, Sagamihara Ku, Kanagawa 2288555, Japan Kitasato Univ Kanagawa Japan 2288555 amihara Ku, Kanagawa 2288555, Japan Kitasato Univ, Sch Med, Dept Pathol, Kanagawa, Japan Kitasato Univ Kanagawa Japan niv, Sch Med, Dept Pathol, Kanagawa, Japan Natl Okura Hosp, Dept Clin Res, Tokyo, Japan Natl Okura Hosp Tokyo Japan atl Okura Hosp, Dept Clin Res, Tokyo, Japan Tokyo Med Univ, Dept Pathol, Tokyo, Japan Tokyo Med Univ Tokyo JapanTokyo Med Univ, Dept Pathol, Tokyo, Japan
Titolo Testata:
DIAGNOSTIC MOLECULAR PATHOLOGY
fascicolo: 4, volume: 10, anno: 2001,
pagine: 265 - 271
SICI:
1052-9551(200112)10:4<265:COTDEM>2.0.ZU;2-P
Fonte:
ISI
Lingua:
ENG
Soggetto:
EPSTEIN-BARR-VIRUS; INSITU HYBRIDIZATION; PCR AMPLIFICATION; CARCINOMA; GENE;
Keywords:
DNA extraction method; paraffin section; beta-globin gene; beta-catenin gene;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
17
Recensione:
Indirizzi per estratti:
Indirizzo: Sato, Y Kitasato Univ, Sch Allied Hlth Sci, Dept Mol Diagnost, Sagamihara Ku, Kanagawa 2288555, Japan Kitasato Univ Kanagawa Japan 2288555 Ku, Kanagawa 2288555, Japan
Citazione:
Y. Sato et al., "Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-embedded tissues", DIAGN MOL P, 10(4), 2001, pp. 265-271

Abstract

To obtain an adequate quality and quantity of DNA from formalin-fixed and paraffin-embedded tissue, six different DNA extraction methods were compared. Four methods used deparaffinization by xylene followed by proteinase K digestion and phenol-chloroform extraction. The temperature of the differentsteps was changed to obtain higher yields and improved quality of extracted DNA. The remaining two methods used microwave heating for deparaffmization. The best DNA extraction method consisted of deparaffinization by microwave irradiation, protein digestion with proteinase K at 48 degreesC overnight, and no further purification steps. By this method. the highest DNA yieldwas obtained and the amplification of a 989-base pair beta -globin gene fragment was achieved. Furthermore, DNA extracted by means of this procedure from five gastric carcinomas was successfully used for single strand conformation polymorphism and direct sequencing assays of the beta -catenin gene. Because the microwave-based DNA extraction method presented here is simple, has a lower contamination risk, and results in a higher yield of DNA compared with the ordinary organic chemical reagent-based extraction method, itis considered applicable to various clinical and basic fields.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/09/20 alle ore 02:59:42