Application of capillary nongel sieving electrophoresis for gene analysis

Citation
Y. Shen et al., Application of capillary nongel sieving electrophoresis for gene analysis, ELECTROPHOR, 20(9), 1999, pp. 1822-1828
Citations number
15
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry & Analysis
Journal title
ELECTROPHORESIS
ISSN journal
0173-0835 → ACNP
Volume
20
Issue
9
Year of publication
1999
Pages
1822 - 1828
Database
ISI
SICI code
0173-0835(199907)20:9<1822:AOCNSE>2.0.ZU;2-U
Abstract
Capillary electrophoresis (CE) has proved to be a strong tool for DNA analy sis and has found abundant applications in the fields of restriction fragme nt sizing, mutation screening, polymerase chain reaction (PCR) product char acterizing and forensic identifying. CE may be the main alternative to slab gel electrophoresis. Capillary nongel electrophoresis is the most favorabl e mode when aiming for this purpose because of its advantages of long lifet ime, easy operation, good reproducibility, and low expense. In this paper, a new kind of sieving matrix, with mannitol as the additive for capillary e lectrophoresis, as well as related methods and their application for gene a nalysis were reported. Nine DNA fragments amplified by multiplex PCR from a normal dystrophin gene were well separated by this system. Three different deletions were found in Duchenne muscular dystrophy (DMD) patients. Three to four copies of the sex-determination region of the Y chromosome (SRY) ge ne, as well as the phenylalanine hydroxylase (PAH) gene, could be detected in mixed samples. The frequencies of short tandem repeats (STR) in PAH gene s was analyzed in 61 normal Chinese individuals and 6 phenylketonuria famil ies. One case of prenatal gene diagnosis was performed. By using this matri x, CE coupled with reverse transcription PCR (RT-PCR), the analysis of the alternative splicing expression pattern of the fragile X mental retardation 1 (FMRI) gene in adult lung tissue was achieved.