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Multicolor karyotype analyses of mouse embryonic stem cells (Guo et.al., 2005)

colcemid (final concentration 0.270 μg/ml) to the culture medium for
2.5 h at 37 ~ C in 5% CO2. Cells were washed with PBS, treated with trypsin-
ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and
centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml
of a prewarmed hypotonic 0.0375 M KC1 solution was added. Cells were
incubated for 20 min at 37 °C. Cells were centrifuged for 5 min at 1200 rpm
and the pellet was resuspended in 3-5 ml of 0.0375 M KC1 solution. The
cells were fixed with methanol/acetic acid (3:1, vohvol) by gently pipetting.
Fixation was performed four times