Chromosome Flow Sorting

Normal Human	Abnormal Human

• Chromosomes can also be analyzed by preparing a monodispersed suspension of chromosomes, staining with fluorochromes and passing them through a flow cytometer. The intensity of each chromosome depends on its DNA content and AT/GC ratio. Chromosomes can also be sorted and chromosome paints can be made by degenerate oligonucleotide-primed (DOP) PCR amplification. These paints can then be hybridized to metaphases or combined to establish SKY kits from any desired species. Chromosome flow sorting of tumors allows reverse painting to study complex chromosomal rearrangements.

• In reverse painting, probes are generated by sorting the rearranged chromosomes from the patient and then hybridizing to normal metaphase spreads. Insight is acquired about the origin of the marker chromosomes and breakpoints involved. Reverse painting also allows a more precise sub-chromosomal assignment for marker chromosomes when chromosomes have been fragmented by multiple translocations and adds information about breakpoints involved in chromosome evolution during tumorigenesis. Phage and cosmid libraries can also be constructed from flow-sorted chromosomes.

Sample Requirements:
Culture requirements:
            Fibroblast – 3 Near Confluent T-150 Flasks with media requirements.
            Lymphoblast – 2-3 Actively growing T-25 Flasks with media requirements.

Please inquire if your sample varies from the listed recommendations.  Please inquire for likely turnaround time.