Two-dimensional DNA typing: A general method for scanning the total human genome
- Aarhus Univ. (Denmark); and others
Two-dimensional (2-D) DNA typing is a method that offers the possibility of analyzing hundreds of loci simultaneously in the human genome. This is done by electrophoretic separation of DNA restriction fragments in two dimensions: firstly according to size, and then in the second dimension by denaturing gradient gel electrophoresis (DGGE) according to base-pair sequence, followed by hybridization analysis using a micro- or minisatellite core probe. Depending on the core probe used in the hybridization analysis, between 150 and 700 restriction fragments are detected as spots in the 2-D DNA typing patterns. Many of the loci corresponding to these spots are polymorphic, and it is anticipated that 10 re-hybridizations with core probes, each detecting 150 informative spots, allow a scanning of the genome at 1500 loci. These properties of 2-D DNA typing make it possible to monitor genetic variability on a total genome scale. In families it is possible to follow the segregation of the 2-D spots, which are inherited in a Mendelian fashion as codominant markers. Due to the high degree of variability in the polymorphic loci, one spot (one allele) is often only represented once in the parents. A spot like that can function as an informative genetic marker without any further knowledge about the other alleles of the locus. We have followed the segregation of several such spots in 3 large CEPH pedigrees, and it has been possible by linkage analysis to map many of these spots. These results show that the spots are indeed inherited in a Mendelian manner, and that they can be scored reliably, and thus underlines the future perspective of 2-D DNA typing as a genetic marker system. An initial database has been set up on the basis of the computerized scanned image of 2-D DNA typings with informations which will mostly concern spot-data from studies on genomic alterations in different cancers, and studies on amplifications of trinucleotide repeats as disease-causing mutations.
- OSTI ID:
- 133468
- Report Number(s):
- CONF-941009--
- Journal Information:
- American Journal of Human Genetics, Journal Name: American Journal of Human Genetics Journal Issue: Suppl.3 Vol. 55; ISSN AJHGAG; ISSN 0002-9297
- Country of Publication:
- United States
- Language:
- English
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