title: Evaluation of the automated fluorescent analysis of the H-ras minisatellite after optimized PCR amplification in comparison with a standardized Southern blot technique.
abstract: Determination of allele sizes? loss of heterozygosity or genetic instability at minisatellite VNTR loci, are routinely performed by the conventional Southern technique. We have investigated the potential use of automated DNA sequencer for the analysis of the H-ras minisatellite. We report the modifications of amplification parameters and electrophoresis conditions on the sequencer. Seventy-one colorectal carcinomas and the corresponding normal tissues were amplified with fluorescent-labeled primers, analyzed on sequencer, and concurrently controlled by Southern blotting. The results on sequencer showed that a Hydrolink matrix used in non-denaturing conditions and a specific analysis software facilitate a more accurate fragment size calculation.
authors.affiliation: CIS BIO INT,GIF SUR YVETTE,FRANCE. CEA,CIS BIO INT,GRENOBLE,FRANCE. CTR JEAN PERRIN,INSERM CRI 9402,MOL ONCOL LAB,F-63011 CLERMONT FERRAN 1,FRANCE.
authors.forename: S
authors.initials: S
authors.lastname: Gosse
authors.forename: S
authors.initials: S
authors.lastname: Sauvaigo
authors.forename: A
authors.initials: A
authors.lastname: Daver
authors.forename: F
authors.initials: F
authors.lastname: Larra
authors.forename: J
authors.initials: J
authors.lastname: Saavedra
authors.forename: J
authors.initials: J
authors.lastname: Marchand
authors.forename: D
authors.initials: D
authors.lastname: Bernardgallon
authors.forename: Y
authors.initials: Y
authors.lastname: Bignon