Science, Vol 238, Issue 4825, 323-329
Copyright © 1987 by American Association for the Advancement of Science
High-speed chromosome sorting
JW Gray,
PN Dean,
JC Fuscoe,
DC Peters,
BJ Trask,
GJ van den Engh,
and
MA Van Dilla
Biomedical Sciences Division, Lawrence Livermore National Laboratory, CA 94550.
Dual-beam high-speed sorting has been developed to facilitate purification of chromosomes based on DNA staining with the fluorescent dyes Hoechst 33258 and chromomycin A3. Approximately 200 chromosomes per second of two types can be sorted from a suspension of chromosomes isolated from human lymphoblasts while fluorescent objects (chromosomes, debris fragments, chromosome clumps, and nuclei) are processed at the rate of about 20,000 per second. This sorting rate is approximately ten times that possible with conventional sorters. Chromosomes of a single type can be sorted with a purity of about 90 percent. DNA from the sorted chromosomes is suitable for construction of recombinant DNA libraries and for gene mapping.
