国際特許
[00290] Karyotype analysis. Karyotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAPS cells were arrested in metaphase by colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37°C in 5% 【C02】.
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 【KC 1】 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; vol/vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides. http://patentscope.wipo.int/search/en/WO2013163296
