Multicolor chromosome banding (MCB) with YAC/BAC-based probes and region-specific microdissection DNA libraries
Multicolor chromosome banding (MCB) allows the delineation of chromosomal regions with a resolution of a few mega base pairs, i.e., slightly below the size of most visible chromosome bands. Based on the hybridization of over lapping region-specific probe libraries, chromosomal subregions are hybridized with probes that fluoresce in distinct wave length intervals, so they can be assigned predefined pseudo-colors during the digital imaging and visualization process. The present study demonstrates how MCB patterns can be produced by region-specific micro dissection derived (mcd) libraries as well as collections of yeast or bacterial artificial chromosomes (YACs and BACs, respectively). We compared the efficiency of an mcd library based approach with the hybridization of collections of locus-specific probes (LSP) for fluorescent banding of three rather differently sized human chromosomes, i.e., chromosomes 2, 13, and 22. The LSP sets were comprised of 107 probes specific for chromosome 2, 82 probes for chromosome 13, and 31 probes for chromosome 22. The results demonstrated a more homogeneous coverage of chromosomes and thus, more desirable banding patterns using the microdissection library-based MCB. This may be related to the observation that chromosomes are difficult to cover completely with YAC and/or BAC clones as single-color fluorescence in situ hybridization (FISH) experiments showed. Mcd libraries, on the other hand, provide high complexity probes that work well as region specific paints, but do not readily allow positioning of break points on genetic or physical maps as required for the positional cloning of genes. Thus, combinations of mcd libraries and locus-specific large insert DNA probes appear to be the most efficient tools for high-resolution cytogenetic analyses.
- Research Organization:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Organization:
- USDOE Director, Office of Science, Office of Biological and Environmental Research; US NIH, (1R01 CA80792, IR33 CA88258); Wilhelm Sander-Stiftung (99.105.1), DAAD, DFG (436 RUS 17/40/00, PO284/6-1),EU (ICA2-CT-2000-10012 and QLRT-1999-31590) (US)
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 815338
- Report Number(s):
- LBNL-51733; R&D Project: 864A1A; TRN: US200319%%175
- Journal Information:
- Cytogenetic and Genome Research, Vol. 97, Issue 1-2; Other Information: Journal Publication Date: 2002; PBD: 23 Jun 2003
- Country of Publication:
- United States
- Language:
- English
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