Generation of five high-complexity painting probe libraries from flow-sorted mouse chromosomes.
Generation of five high-complexity painting probe libraries from flow-sorted mouse chromosomes.
复制标题
从流式分选的小鼠染色体生成五个高复杂性绘画探针库。
DOI：
10.1006/geno.1994.1326
复制
发表时间：
1994
期刊：
影响因子：
4.4
通讯作者：
Gray,JW
中科院分区：
文献类型：
--
作者：
Weier,HU;Polikoff,D;Fawcett,JJ;Greulich,KM;Lee,KH;Cram,S;Chapman,VM;Gray,JW
Mouse metaphase chromosomes were purified by flow sorting from the murine fibroblast cell lineMus 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-directedin 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 theM. 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.
