2.8 Metaphase arrest and chromosome fusion studies
Cells were seeded in 6 well plates followed by 1 µg/ml of colcemid was
added and incubated for 3 hours at 37°C with 5% CO2. Cells were
collected by using trypsinization protocol, centrifuged at 1000rpm for 4
mins at RT, resuspended the pellet with 0.56% ice-cold KCL solution,
mixed gently and stand for 6 min at room temperature. Drop-by-drop of
5ml fixative solution 3:1 ratio (Methanol: Acetic acid) was added to the
cells. After centrifuge, 1 ml of fixative mix and small volume of
mixture was released dropwise on a clean glass slide. Mounting medium
with DAPI (Nuclear stain) was added to the glass slide and coverslip was
placed and mount with nail polish for observation and documentation
under EVOS epi-fluorescence microscope.