Protocol by Ross Francis
Modified by Sudhir Nayak
To visualize DNA, incubate fixed samples with 100 ng/ml 4′,6′-diamidino-2-phenylindole hydrochloride (DAPI) in PBT. The staining time depends on the type of sample and varies from 5-30 min. For example, 5 min is long enough for dissected gonad arms but longer incubation (20-30 min) is required for whole mount animals(*). After staining rinse 2x with PBT and mount in DABCO.
*For whole mount animals use methanol fixation.
Materials
PBS: Sambrook et al. (Molecular Cloning).
PBT: PBS containing 0.1% Tween 20.
DAPI: 100 ug/ml stock solution in dH2O. Store at -20C.
DABCO — 1% 1,4-diazobicyclo[2,2,2]-octane (DABCO) in 90% glycerol in PBS. Store at -20 good for years.