Multilocus genomic mapping with intracisternal A-particle proviral oligonucleotide probes hybridized to mouse DNA in dried agarose gels

Recently, oligonucleotide probes that detect intracisternal A-particle (IAP) gene subfamilies with a limited number of proviral copies have been shown to be useful multilocus markers. A procedure for hybridization of these probes has been developed and is described. In summary, the main features of...

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Bibliographic Details
Published inElectrophoresis Vol. 16; no. 2; p. 179
Main Author Lueders, K K
Format Journal Article
LanguageEnglish
Published Germany 1995
Subjects
Animals
Base Sequence
Chromosome Mapping - methods
DNA - chemistry
DNA - genetics
DNA Transposable Elements
Electrophoresis, Agar Gel - methods
Genes, Intracisternal A-Particle
Genetic Markers
Methylation
Mice
Mice, Inbred Strains
Molecular Sequence Data
Mutation
Nucleic Acid Hybridization
Oligonucleotide Probes - genetics
Proviruses - genetics
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Summary:Recently, oligonucleotide probes that detect intracisternal A-particle (IAP) gene subfamilies with a limited number of proviral copies have been shown to be useful multilocus markers. A procedure for hybridization of these probes has been developed and is described. In summary, the main features of the method are the following: (i) A pulse controller is used during agarose gel electrophoresis to improve resolution of restriction fragments in genomic DNA. (ii) Hybridization is performed in a dried gel. (iii) The hybridized gel is washed in tetramethylammonium chloride to eliminate differences in oligonucleotide composition on hybrid stability. Use of the procedure is demonstrated by genomic mapping of IAP loci in the AXB BXA recombinant inbred mouse strains, identification of hypomethylated loci in tumor cells, and detection of a transposed IAP provirus previously identified as the basis for a mutation at the agouti locus.
ISSN:0173-0835
DOI:10.1002/elps.1150160132