A reliable preparation of mon-dispersed chromosome suspensions for flow cytometry.
A reproducible, simple and rapid protocol was developed to prepare mono-dispersed chromosome suspensions for flow cytometric analysis. The procedure basically employes: 1. twice rinsing of monolayer cultures to eliminate dead cells and cellular debris, 2. mild fixation of mitotic cells with 1% acetic acid, and 3. ultra sonic treatment to release single metaphase chromosomes. The procedure takes less than 30 min. Isolated chromosomes are storable over months at 4 degrees C. Despite mild fixation many biochemical studies and experiments applied on such fixed chromosomes purified and enriched using electronic sorting are feasible: 1. gene and restriction mapping, 2. cloning of specific gene sequences, and 3. gene frequency analysis.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002478:Cells, Cultured; D002875:Chromosomes / Q000032:analysis*; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry; D008647:Mesocricetus; D008938:Mitosis; D014465:Ultrasonics
D004247:DNA