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Titolo:
A study of parameters that influence the HPLC and CE separation of double stranded DNA fragments and DNA mutants
Autore:
Issaq, HJ; Xu, HY; Chan, KC;
Indirizzi:
NCI, SAIC Frederick, Frederick, MD 21702 USA NCI Frederick MD USA 21702NCI, SAIC Frederick, Frederick, MD 21702 USA
Titolo Testata:
JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES
fascicolo: 16, volume: 24, anno: 2001,
pagine: 2381 - 2398
SICI:
1082-6076(2001)24:16<2381:ASOPTI>2.0.ZU;2-T
Fonte:
ISI
Lingua:
ENG
Soggetto:
CONFORMATION POLYMORPHISM ANALYSIS; SINGLE BASE SUBSTITUTIONS; POLYMERASE CHAIN-REACTION; CAPILLARY-ELECTROPHORESIS; POINT MUTATIONS; LIQUID-CHROMATOGRAPHY; TEMPERATURE; MISMATCHES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Physical, Chemical & Earth Sciences
Citazioni:
23
Recensione:
Indirizzi per estratti:
Indirizzo: Issaq, HJ NCI, SAIC Frederick, POB B, Frederick, MD 21702 USA NCI POB B Frederick MD USA 21702 POB B, Frederick, MD 21702 USA
Citazione:
H.J. Issaq et al., "A study of parameters that influence the HPLC and CE separation of double stranded DNA fragments and DNA mutants", J LIQ CHR R, 24(16), 2001, pp. 2381-2398

Abstract

The present study evaluates HPLC and CE experimental parameters, such as column packing properties, mobile phase pH, column temperature, and gel typeon the separation of DNA fragments and heteroduplexes. The results of thisstudy show that both HPLC and CE are useful techniques for the separation of DNA fragments and for the detection of DNA mutants. Not all HPLC columns tested resolved the DNA fragments or detected the mutation. The packing material type and physical and chemical properties play a significant role. It was found that ion-pair DHPLC is easier and faster than single capillary CE for detecting DNA mutants at their melting temperature (5 min. vs 25 min.). However, CE is faster for the separation of DNA fragments (6.5 min. vs 20 min.). Most HPLC column packing materials tested resolved small DNA fragments, less than 200 bp, but not large fragments, greater than 200 bp. DNA fragments, 21-587 bp, are resolved by DHPLC at 40-50 degreesC, but not at room temperature, using special columns; ultra pure, large pore, high coverage C-18-silica, nonporous C-14-silica or nonporous polystyrene/divinylbenzene and ion-pair gradient elution. CE can resolve the 21-587 bp fragments at room temperature, and at 30 to 50 degreesC. It was also found that fluorocarbon coated capillary filled with polyacryl amide or hydroxycellulose liquid polymers, was more stable thanDB-17, however, both capillaries gave good and reproducible CE results.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/01/20 alle ore 06:30:10