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Titolo:
Multitemperature single-strand conformation polymorphism
Autore:
Kaczanowski, R; Trzeciak, L; Kucharczyk, K;
Indirizzi:
Kucharczyk TE Co, PL-02495 Warsaw, Poland Kucharczyk TE Co Warsaw PolandPL-02495 k TE Co, PL-02495 Warsaw, Poland Int Inst Mol & Cellular Biol, Dept Mol Biol, Warsaw, Poland Int Inst Mol &Cellular Biol Warsaw Poland ept Mol Biol, Warsaw, Poland M Nencki Inst Expt Biol, PL-02093 Warsaw, Poland M Nencki Inst Expt Biol Warsaw Poland PL-02093 , PL-02093 Warsaw, Poland
Titolo Testata:
ELECTROPHORESIS
fascicolo: 16, volume: 22, anno: 2001,
pagine: 3539 - 3545
SICI:
0173-0835(200110)22:16<3539:MSCP>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
TUMOR-SUPPRESSOR GENE; POINT MUTATIONS; P53 GENE;
Keywords:
single-strand conformation polymorphism single-stranded DNA conformation; p53 gene; single nucleotide polymorphism; mutation detection; genomics;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
19
Recensione:
Indirizzi per estratti:
Indirizzo: Kucharczyk, K Kucharczyk TE Co, Ul Dzieci Warszawy 31-20, PL-02495 Warsaw,Poland Kucharczyk TE Co Ul Dzieci Warszawy 31-20 Warsaw Poland PL-02495
Citazione:
R. Kaczanowski et al., "Multitemperature single-strand conformation polymorphism", ELECTROPHOR, 22(16), 2001, pp. 3539-3545

Abstract

Changes of gel temperature during single-strand conformation polymorphism (SSCP) electrophoresis increase the sensitivity of mutation detection in polymerase chain reaction (PCR) products and significantly reduce the overalltime and costs of analysis. Based on these findings, a new method for single nucleotide polymorphism (SNP) and point mutation detection - multitemperature single-strand conformation polymorphism (MSSCP) was devised. In orderto control the gel temperature with 0.1 degreesC accuracy during electrophoresis, new equipment was developed. We demonstrated that increasing the gel temperature by 8 degreesC or decreasing it by 10 degrees from 23 degreesCled to the disappearance of all electrophoretic differences between five alleles of exon 8 of the human p53 gene during the SSCP analysis. The interesting result was the detection of two additional SNPs (out of seven analyzed) in exon 7 of the human PAH gene during a one hour MSSCP electrophoresis. This result is better than that obtained by three classical SSCP analyses of the same samples at different but constant gel temperatures. We advocatethe MSSCP technology as a fast, reliable, and cost-effective tool for the screening and preselection stage of genomics surveys, especially when a high variability of the analyzed DNA fragment is expected.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 25/01/20 alle ore 03:11:03