Rapid generation of whole chromosome painting probes (WCPs) by chromosome microdissection

Journal Article · Fri Jul 01 04:00:00 EDT 1994 · Genomics

DOI:https://doi.org/10.1006/geno.1994.1350· OSTI ID:255177

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

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1. National Institutes of Health, Bethesda, MD (United States)

A strategy for rapid construction of whole chromosome painting probes (WCPs) by chromosome microdissection has recently been developed. WCPs were prepared from 20 copies of each target chromosome microdissected from normal human metaphase chromosomes and then directly amplified by PCR using a universal primer. Fifteen WCPs, including chromosomes 1, 3, 6, 7, 9, 12, 13, 14, 15, 17, 19, 20, 21, 22, and X, have been generated using this strategy. The probe complexity and hybridization specificity of these WCPs have been characterized by gel electrophoresis and fluorescence in situ hybridization. Analysis of WCPs constructed by chromosome microdissection indicated that microdissected WCPs invariably provide strong and uniform signal intensity with no cytologically apparent cross-hybridization. To demonstrate the application of WCPs generated from microdissection, the authors have used these probes to detect complex chromosome rearrangements in a melanoma cell line, UM93-007. Two different translocations involving three chromosomes [t(1;3;13) and t(1;7;13)] have been identified, both of which were undetectable by conventional banding analysis. Further application of these WCPs (including generation of WCPs from mouse and other species) should greatly facilitate the cytogenetic analysis of complex chromosome rearrangements. 35 refs., 4 figs.

• OSTI ID: 255177
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 1 Vol. 22; ISSN GNMCEP; ISSN 0888-7543
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
CHROMOSOMAL ABERRATIONS
DETECTION
DNA HYBRIDIZATION
HUMAN CHROMOSOME 1
HUMAN CHROMOSOME 12
HUMAN CHROMOSOME 13
HUMAN CHROMOSOME 14
HUMAN CHROMOSOME 15
HUMAN CHROMOSOME 17
HUMAN CHROMOSOME 19
HUMAN CHROMOSOME 21
HUMAN CHROMOSOME 22
HUMAN CHROMOSOME 3
HUMAN CHROMOSOME 6
HUMAN CHROMOSOME 7
HUMAN CHROMOSOME 9
HUMAN X CHROMOSOME