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CHROMOSOMAL BAR CODES PRODUCED BY MULTICOLOR FLUORESCENCE INSITU HYBRIDIZATION WITH MULTIPLE YAC CLONES AND WHOLE CHROMOSOME PAINTING PROBES
Author(s): LENGAUER C, SPEICHER MR, POPP S, JAUCH A, TANIWAKI M, NAGARAJA R, RIETHMAN HC, DONISKELLER H, DURSO M, SCHLESSINGER D, CREMER T
Source: HUMAN MOLECULAR GENETICS    Volume: 2    Issue: 5    Pages: 505-512    Published: MAY 1993  
Times Cited: 79     References: 48     
Abstract: Colored chromosome staining patterns, termed chromosomal 'bar codes' (CBCs), were obtained on human chromosomes by fluorescence in situ hybridization (FISH) with pools of Alu-PCR products from YAC clones containing human DNA inserts ranging from 100 kbp to 1 Mbp. In contrast to conventional G- or R-bands, the chromosomal position, extent, individual color and relative signal intensity of each 'bar' could be modified depending on probe selection and labeling procedures. Alu-PCR amplification products were generated from 31 YAC clones which mapped to 37 different chromosome bands. For multiple color FISH, Alu-PCR amplification products from various clones were either biotinylated or labeled with digoxigenin. Probes from up to twenty YAC clones were used simultaneously to produce CBCs on selected human chromosomes. Evaluation using a cooled CCD camera and digital image analysis confirmed the high reproducibility of the bars from one metaphase spread to another. Combinatorial FISH with mixtures of whole chromosome paint probes was applied to paint seven chromosomes simultaneously in different colors along with a set of YAC clones which map to these chromosomes. We discuss the potential to construct analytical chromosomal bar codes adapted to particular needs of cytogenetic investigations and automated image analysis.
Document Type: Article
Language: English
Addresses:
1. INST HUMAN GENET & ANTHROPOL, IM NEUENHEIMER FELD 328, W-6900 HEIDELBERG, GERMANY
2. WASHINGTON UNIV, SCH MED, DEPT MOLEC MICROBIOL, ST LOUIS, MO 63110 USA
3. WISTAR INST ANAT & BIOL, PHILADELPHIA, PA 19104 USA
4. WASHINGTON UNIV, SCH MED, DEPT GENET, ST LOUIS, MO 63110 USA
5. IST INT GENET & BIOFIS, I-80125 NAPLES, ITALY
Publisher: OXFORD UNIV PRESS UNITED KINGDOM, WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP
Subject Category: Biochemistry & Molecular Biology; Genetics & Heredity
IDS Number: LA746
ISSN: 0964-6906
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