Rapid generation of region-specific probes by chromosome microdissection: Application to the identification of chromosomal rearrangements

Conference · Fri Jan 01 04:00:00 EST 1993 · Cytometry (Baltimore); (United States)

OSTI ID:6572957

Trent, J M; Guan, X Y; Zang, J; Meltzer, P S ^[1]

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1. Univ. of Michigan, Ann Arbor (United States)

The authors present results using a novel strategy for chromosome microdissection and direct in vitro amplification of specific chromosomal regions, to identify cryptic chromosome alterations, and to rapidly generate region-specific genomic probes. First, banded chromosomes are microdissected and directly PCR amplified by a procedure which eliminates microchemistry (Meltzer, et al., Nature Genetics, 1:24, 1992). The resulting PCR product can be used for several applications including direct labeling for fluorescent in situ hybridization (FISH) to normal metaphase chromosomes. A second application of this procedure is the extremely rapid generation of chromosome region-specific probes. This approach has been successfully used to determine the derivation of chromosome segments unidentifiable by standard chromosome banding analysis. In selected instances these probes have also been used on interphase nuclei and provides the potential for assessing chromosome abnormalities in a variety of cell lineages. The microdissection probes (which can be generated in <24 hours) have also been utilized in direct library screening and provide the possibility of acquiring a significant number of region-specific probes for any chromosome band. This procedure extends the limits of conventional cytogenetic analysis by providing an extremely rapid source of numerous band-specific probes, and by enabling the direct analysis of essentially any unknown chromosome region.

• OSTI ID: 6572957
• Report Number(s): CONF-9303114--
• Conference Information: Journal Name: Cytometry (Baltimore); (United States) Journal Volume: 6
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMPLIFICATION
CHROMOSOMAL ABERRATIONS
CYTOLOGICAL TECHNIQUES
DETECTION
DNA
DNA HYBRIDIZATION
FLUORESCENCE
HYBRIDIZATION
LIBRARIES
LUMINESCENCE
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PROBES
PRODUCTION
SCREENING