NEW APPROACHES TO FLOW SORTING OF CHROMOSOMES

PURIFICATION OF INDIVIDUAL CHROMOSOMES BY FLOW SORTING IS OFTEN A FIRST STEP IN DEFINING THE STRUCTURE OF NORMAL AND ABNORMAL HUMAN GENES. THE POLYMERASE CHAIN REACTION (PCR) DRASTICALLY REDUCES THE AMOUNT OF SAMPLE REQUIRED FOR ISOLATION OF CHROMOSOMAL DNA, MAKING IT FEASIBLE TO DEVELOP SIMPLER, LESS EXPENSIVE CHROMOSOME SORTERS. THE PERFORMANCE OF AN INSTRUMENT USING AIR-COOLED ULTRAVIOLET (UV) AND BLUE HELIUM-CADMIUM (HE-CD) LASER LIGHT SOURCES IN FLOW CYTOMETRY OF CHROMOSOMES STAINED WITH DAPL/OLIVOMYCIN OR HOECHST 33258/CHROMOMYCIN A3 WILL BE EVALUATED TO DETERMINE WHETHER THE SYSTEM CAN RESOLVE THE HUMAN KARYOTYPE AS WELL AS CAN EXISTING COMMERCIAL APPARATUS. AS AN ALTERNATIVE, SEVERAL NUCLEIC ACID FLUOROCHROMES NOT PREVIOUSLY APPLIED TO CHROMOSOME ANALYSIS AND POTENTIALLY USABLE IN A LARGE NUMBER OF EXISTING FLOW SORTERS WILL BE EXAMINED AS ADJUNCTS OR ALTERNATIVES TO STAINS PRESENTLY USED FOR BIVARIATE FLOW KARYOTYPING.