Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
TEMPERATURE-PROGRAMMED CAPILLARY ELECTROPHORESIS FOR THE ANALYSIS OF HIGH-MELTING POINT MUTANTS IN THALASSEMIAS
Autore:
GELFI C; RIGHETTI PG; TRAVI M; FATTORE S;
Indirizzi:
LITA,VIA FRATELLI CERVI 93,SEGRATE 20090 MILAN ITALY CNR,IST TECNOL BIOMED AVANZATE I-20131 MILAN ITALY UNIV VERONA,DEPT AGR & IND BIOTECHNOL I-37100 VERONA ITALY ICP,LAB RIC CLIN MILAN ITALY
Titolo Testata:
Electrophoresis
fascicolo: 5, volume: 18, anno: 1997,
pagine: 724 - 731
SICI:
0173-0835(1997)18:5<724:TCEFTA>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
GRADIENT GEL-ELECTROPHORESIS; PRENATAL-DIAGNOSIS; POLYMER-SOLUTIONS; BETA-THALASSEMIA; MOLECULAR-BASIS; DNA; SEPARATION; FRAGMENTS; MUTATIONS; ARRAY;
Keywords:
N-SUBSTITUTED ACRYLAMIDES; DNA ELECTROPHORESIS; CAPILLARY ELECTROPHORESIS; THALASSEMIAS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
35
Recensione:
Indirizzi per estratti:
Citazione:
C. Gelfi et al., "TEMPERATURE-PROGRAMMED CAPILLARY ELECTROPHORESIS FOR THE ANALYSIS OF HIGH-MELTING POINT MUTANTS IN THALASSEMIAS", Electrophoresis, 18(5), 1997, pp. 724-731

Abstract

The behavior of different sieving polymers for unambiguous determination of point mutations in genomic DNA, based on electrophoresis in thin capillaries, is evaluated. High melters from thalassemia patients are separated by exploiting the principle of denaturing gradient gel electrophoresis, in fact, of its variant utilizing temperature gradients (TGGE), along the migration path, encompassing the melting points of both homo- and heteroduplex, polymerase chain reaction (PCR)-amplified DNA fragments. Unlike TGGE, where the temperature gradient exists along the separation space, the denaturing temperature gradient in the fused-silica capillaries is time-programmed, so as to reach the T-m's of all species under analysis prior to electrophoretic transport past thedetector window. The DNA fragments are injected in a capillary maintained (by combined chemical and thermal means) just below the expected T, values. The Delta T applied is rather minute (1-1.5 degrees C) and the temperature gradient quite shallow (e.g., 0.05 degrees C/min). Thedenaturing thermal gradient is generated internally, via Joule heat produced by voltage ramps. This method is applied to the analysis of the most common point mutations in thalassemias, characterized by being high melters (in the temperature range of 60-62 degrees C) in presenceof 6 M urea. Point mutants are fully resolved into a spectrum of fourbands only when poly(N-acryloylaminopropanol) and hydroxfethylcellulose are used. However. the former offers the best separation capabilityat such high temperatures.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/11/20 alle ore 12:30:30