70% Ethanol at – 20oC
DNA Staining Buffer:
Triton–x 100 0.75ml
Propidium iodide 0.025g
Ribonuclease A 0.005g
Distilled water 250 ml
cells from each sample into a polypropylene tube and centrifuge at 250 x g for 5
supernatant as completely as possible without disturbing the pellet and add
1 ml of –20oC 70% EtOH dropwise to the cell pellet while
Keep cells at
-20oC until the day of DNA staining (cells can be stored for several
weeks at -20oC).
On the day of
DNA staining, take samples out of the freezer and spin them down by
centrifugation at 250 x g for 5 min. Remove
the supernatant as completely as possible without disturbing the cell pellet.
Add 1 ml of
DNA staining buffer to the cell pellet and vortex gently and briefly.
Keep cells for 15 min in the staining solution before acquisition on the
Cat# C7254 Sigma, St. Louis, MO
Triton-x 100 Cat# x-100 "
Ribonuclease A Cat# R4875 "
Propidium iodide Cat# 537059 Calbiochem, San Diego, CA