Comparative Molecular Cytogenetics Core Facility
Spectral Karyotyping (SKY)
SKY is a 24-color, whole-chromosome painting assay that allows the simultaneous
visualization of each chromosome in different colors. Five spectrally distinct dyes
are used in combination to create a cocktail of probes unique to each chromosome.
The probe mixture is hybridized to metaphase chromosomes on a slide and then visualized
with a spectral cube, which allows the measurement of the entire emission spectrum
with a single exposure. The image is processed by computer software that can distinguish
differences in color not discernible to the naked eye by assigning a numerical value
to the RGB and pseudo color images.
SKY can detect chromosomal material of unknown origin, complex rearrangements, translocations,
large deletions, duplications, aneuploidy, and more. Some disadvantages of SKY include
ineffective detection of microdeletions and inversions. It can only be performed
on dividing cells.
Investigator responsibilities:
- Provide fixed, frozen, or actively growing cells.
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Testing Options
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Identifies chromosomal aberrations
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Translocations (balanced)
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Yes
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Translocations (unbalanced)
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Yes
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Copy number changes
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Yes
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Additions (>10MB)
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Yes
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Deletions (>10MB)
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Yes
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Deletions of specific genes/loci
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No
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Inversions
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Sometimes
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Insertions
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Yes
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Identifies presence of markers
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Yes
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Distinguishes between double minutes and homogeneously staining regions
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Yes
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Identifies chromosomal origins
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Yes
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Identifies specific p/q arms/bands
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No
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Requires specifically labeled probes
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Yes
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More than two probes at one time
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Yes
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Requires viable material
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Yes
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Requires metaphase spreads
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Yes
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Applicable to interphase nuclei
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No
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Applicable to DNA extracted from paraffin embedded tissue
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No
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Affected by normal tissue contamination
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Yes
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Identifies tumor heterogeneity
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Yes
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Applicable for studies in other species
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Yes
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Sample Requirements:
- Peripheral blood: 5-10 mls in Heparin container
- Culture: Culture must be actively growing with media supplied. A T-75 flask
is preferred. Tissue culture flask must not be vented. This prevents spillage and
contamination during transit. Do not fill beyond recommendation of flask i.e. (to
the absolute top of flask or tube).
Please inquire if your sample varies from the listed recommendations. Please inquire
for likely turnaround time.