Yeast Nuclear Staining

DAPI nuclear staining

Materials

Method

  • Harverst 1 ml of yeast culture at 5⋅106 cell/ml;
  • Fix them with 2 ml EtOH 100% for at least 30 minutes;
  • Wash the cells twice with 2 ml 1X PBS;
  • Resuspend in 1 ml 1X PBS;
  • Cells can be stored at 4°C for a few weeks;
  • Centrifuge the cells and resuspend them in 50 μl of 0.2 μg/ml DAPI(DAPI stock solution 1 mg/ml is 5000X) and leave them 30' in a dark place;
  • Wash two times the cells with H2O;
  • After the addition of the second wash sonicate the cells 4-5 seconds;
  • Centrifuge 5' at 5000 rpm;
  • Resuspend in 30 μl of 50% glycerol and look ~1µl of cells under the microscope;