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.