Replication kinetics of Chinese hamster chromosomes as revealed by bivariate flow karyotyping
✍ Scribed by C. Cremer; J. W. Gray
- Publisher
- John Wiley and Sons
- Year
- 1983
- Tongue
- English
- Weight
- 519 KB
- Volume
- 3
- Category
- Article
- ISSN
- 0196-4763
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✦ Synopsis
In this paper we show that the replication kinetics of individual chromosomes can be estimated by bivariate flow cytometric analysis of chromosomes isolated from 5-bromo-Z'-deoxyuridine (BrdUrd) treated cells. To study the timing of replication of chromosomes, Chinese hamster M3-1 cells were grown for different times in BrdUrd containing medium. Colcemid was added during the last 90 minutes of each BrdUrd labeling period. Chromosomes were isolated from the Colcemid blocked mitotic cells and stained with Hoechst 33258 (HO) and chromomycin A3 (CA3). Bivariate HO-CA3 flow karyotypes were measured in a dual beam flow cytometer. Labeling cells with BrdUrd for 2, 2%, 3, 3%, and 4 hours before chromosome isolation resulted in progressive quenching of the HO fluorescence of chromosomes Y and 10, 11, M2 while the HO fluorescence of