title: DNA probes for FISH analysis of C-negative regions in human chromosomes.
abstract: Fluorescent in situ hybridization (FISH) is a powerful technology for studying the chromosome organization and aberrations as well as for searching the homology between chromosomal regions in mammals. Currently, FISH is used as a simple, rapid, and reliable technique for analyzing chromosomal rearrangements and assigning chromosomal breakpoints in modern diagnosing of chromosomal pathology. In addition to cloned DNA fragments, the DNA probes produced by sequence-independent polymerase chain reaction are widely used in FISH assays. As a rule, the DNA probes generated from a genomic or chromosomal DNA by whole genome amplification are enriched for repetitive elements and, consequently, efficient FISH analysis requires that repetitive DNA hybridization is suppressed. The linker-adapter polymerase chain reaction (LA-PCR) using the genomic DNA hydrolyzed with HaeIII and RsaI restriction endonucleases allows the repetitive DNA fraction in DNA probe to be decreased and gene-rich DNA to be predominantly amplified. The protocol described here was proposed for production of the DNA probes for enhanced analysis of the C-negative regions in human chromosomes.
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russian Federation.
authors.forename: Evgeniy S
authors.initials: ES
authors.lastname: Morozkin
authors.forename: Tatyana V
authors.initials: TV
authors.lastname: Karamysheva
authors.forename: Pavel P
authors.initials: PP
authors.lastname: Laktionov
authors.forename: Valentin V
authors.initials: VV
authors.lastname: Vlassov
authors.forename: Nikolay B
authors.initials: NB
authors.lastname: Rubtsov
mesh_terms: D002478:Cells, Cultured; D002877:Chromosomes, Human / Q000235:genetics*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry; D005819:Genetic Markers; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016133:Polymerase Chain Reaction; D012235:Rhodamines / Q000737:chemistry
chemical_list: C114557:5-carboxytetramethylrhodamine succinimidyl ester; D015342:DNA Probes; D005456:Fluorescent Dyes; D005819:Genetic Markers; D012235:Rhodamines