Purifying human Y chromosomes by flow cytometry and sorting.
A method of producing an enriched sample of human Y chromosomes from peripheral blood lymphocytes is described. Metaphase chromosomes were prepared from peripheral blood lymphocytes donated by 17 normal male individuals. A suspension of chromosomes in a polyamine buffer was produced from each sample, stained with the fluorescent dye Hoechst 33258, and passed through a flow cytometer and sorter. Following analysis of the 17 fluorescence distributions, a single donor was found giving a separate peak corresponding to the Y chromosome. Seventy percent of the chromosomes sorted from this peak were identified as Y chromosomes. Batches of a million Y chromosomes were produced from each of several 40 ml donations of peripheral blood. These were assessed for the amount of Y DNA present and used to construct a DNA library.
D006690:Bisbenzimidazole; D002458:Cell Fractionation / Q000379:methods*; D002871:Chromosome Banding; D005434:Flow Cytometry; D006801:Humans; D008214:Lymphocytes / Q000648:ultrastructure; D008297:Male; D014998:Y Chromosome
D006690:Bisbenzimidazole