Generation of five high-complexity painting probe libraries from flow-sorted mouse chromosomes
- Univ. of California, San Francisco, CA (United States)
- Lawrence Berkeley Lab., CA (United States); and others
Mouse metaphase chromosomes were purified by flow sorting from the murine fibroblast cell line Mus spretus clone 5A. The authors sorted chromosomes that fell into five individual peaks based on the Hoechst 33258/chromomycin A3 DNA histogram: three peaks corresponding to the least amount of DNA and two peaks representing chromosomes with the most DNA content. This is the first example of the successful application of bivariate flow karyotyping to murine chromosome sorting. They then applied primer-directed in vitro DNA amplification using the polymerase chain reaction (PCR) to generate and label larger amounts of chromosome-specific DNA. In situ hybridization showed specific binding of the PCR products to mouse chromosomes Y, 19, 18, 3, and X as well as chromosomes 1 and 2. The combination of chromosome sorting from the M. spretus cell line and PCR proved to be highly valuable for generation of pools of DNA fragments that exhibit specific binding to mouse chromosomes and can be used to identify and delineate mouse metaphase chromosomes. 13 refs., 3 figs.
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 250058
- Journal Information:
- Genomics, Vol. 21, Issue 3; Other Information: PBD: Jun 1994
- Country of Publication:
- United States
- Language:
- English
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