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Titolo:
Conditions for single-strand conformation polymorphism (SSCP) analysis of BRCA1 gene using an automated electrophoresis unit
Autore:
Campos, B; Diez, O; Cortes, J; Domenech, M; Pericay, C; Alonso, C; Baiget, M;
Indirizzi:
Hosp Santa Creu & Sant Pau, Serv Genet, Barcelona 08025, Spain Hosp Santa Creu & Sant Pau Barcelona Spain 08025 Barcelona 08025, Spain Hosp Santa Creu & Sant Pau, Med Oncol Serv, Barcelona, Spain Hosp Santa Creu & Sant Pau Barcelona Spain Oncol Serv, Barcelona, Spain
Titolo Testata:
CLINICAL CHEMISTRY AND LABORATORY MEDICINE
fascicolo: 5, volume: 39, anno: 2001,
pagine: 401 - 404
SICI:
1434-6621(200105)39:5<401:CFSCP(>2.0.ZU;2-0
Fonte:
ISI
Lingua:
ENG
Soggetto:
POINT MUTATIONS; OVARIAN-CANCER; OPTIMIZATION; BREAST;
Keywords:
BRCA1 gene; SSCP automated analysis; breast cancer;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
9
Recensione:
Indirizzi per estratti:
Indirizzo: Diez, O Hosp Santa Creu & Sant Pau, Serv Genet, Pare Claret 167, Barcelona08025, Spain Hosp Santa Creu & Sant Pau Pare Claret 167 Barcelona Spain 08025
Citazione:
B. Campos et al., "Conditions for single-strand conformation polymorphism (SSCP) analysis of BRCA1 gene using an automated electrophoresis unit", CLIN CH L M, 39(5), 2001, pp. 401-404

Abstract

The single-strand conformation polymorphism procedure has been applied in routine testing for hereditary diseases and cancer. However, temperature, running time, gel composition, fragment length, etc. can influence its sensitivity. Mutation detection in the clinical setting depends on the development of automated technology, especially for large genes such as the breast cancer gene BRCA1. We analysed DNA samples with BRCA1 mutations in an automated system (GenePhor System; Amersham-Pharmacia Biotech, Uppsala, Sweden). The concentrations of DNA template and PCR primers, the effect of chilling after denaturation, and the temperature and time of the electrophoresis were investigated. All band-shifts were detected by electrophoresis at 5 degreesC for 2 h 15 min. Concentrations of DNA and samples used in the PCR did not affect the SSCP pattern, but chilling the PCR product in ice after denaturation was required. The type and position of mutation in the fragments did not influence the probability of a mobility shift, although SSCP analysiswas more sensitive for fragments shorter than 350 bp. This automated SSCP method meets the requirements of fast turnaround and sensitivity and can bereadily adapted to the screening of large genes such as BRCA1.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 25/01/20 alle ore 06:17:05