Division of Animal Genetics, Department of Animal Science and Animal Health, The Royal Veterinary and Agricultural University, Bülowsvej 13, DK-1870 Frederiksberg C, Denmark
Some of our mink cosmids contain strong centromere repeats. When they are used for in situ hybridization on sperm cells they will reveal non disjunction events. The pictures below show results from such experiments. Before in situ hybridization the slides had been pre treated with dithiotreitol (DTT) for 1-2 minutes (200 mM KOH, 50 mM DTT) and neutralized with (900mM Tris-HCl, pH 8.3, +300mM KCl, 200 mM HCl) and finally washed with distil water.
In situ hybridization with cosmid AG64 which is located on chromosome 8, the cell in the insert show nondisjunction as there are two dots.
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In situ hybridization with cosmid AG63 which is located on chromosome 9, above left.
In situ hybridization with cosmid AG32 which is located on chromosome 11, above right.
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In situ hybridization with cosmid AG34, above left (nucleolar organizer)which is located on
chromosome 2 and 5, two dots are expected on each sperm cell.
In situ hybridization with cosmid AG25, above right, which is located on Y chromosome, half the cell
which are Y chromosome carring should show signal. The signals are somewhat diffuse.
For more details see here
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