Comparative Genomic Hybridization onto Dense Arrays of DNA Clones: Development and Application to Breast Cancer Genomes.
Abstract
The amplification of chromosome 20q occurs in 15-20% of primary breast tumors and correlates with poor prognosis indicating that this region may contains gene(s) involved in breast cancer development. In order to define this region with high precision, we are developing methods to perform comparative genomic hybridization to dense clone arrays. Tumor and reference DNAs are labeled with different fluorochromes and simultaneously hybridized to an immobilized set of mapped clones spanning the length of chromosome 20. Copy number abnormalities involving a clone are indicated by changes in the tumor/normal fluorescence ratio compared to average. By titrating X DNA probe i n a background of human genomic DNA, we show that this method possesses sufficient sensitivity to detect single copy deletions as well as amplification, and is linear over several orders of magnitude. Using manually prepared 30-60 clones arrays, breast cancer cell lines and primary tumors reveal that 20q is suprisingly complex, possesing at least five regions of amplification and possibly one deletion. Finally, an automated printer is being developed to prepare large numbers (%100) of arrays in parallel of higher (up to 1 0% spots/cm2) density.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1997
- Accession Number
- ADA334089
Entities
People
- Daniel Pinkel
- Steven M. Clark
Organizations
- University of California, San Francisco