Chromosome Flow Sorting
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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.
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