| Autor: |
Weier, Heinz-Ulrich G., Polikoff, Daniel, Fawcett, John J., Greulich, Karin M., Lee, Kwang-Ho, Cram, Scott, Chapman, Verne M., Gray, Joe W. |
| Zdroj: |
Genomics; June 1994, Vol. 21 Issue: 3 p641-644, 4p |
| Abstrakt: |
Mouse metaphase chromosomes were purified by flow sorting from the murine fibroblast cell line Mus spretusclone 5A. We 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. We then applied primer-directed in vitroDNA amplification using the polymerase chain reaction (PCR) to generate and label larger amounts of chromosome-specific DNA. In situhybridization 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. spretuscell 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. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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