User talk:Chenisr

'DAPI staining'

DAPI-dihydrochloride (D9542 Sigma, 4′,6-Diamidino-2-phenylindole dihydrochloride ) Stock solution preparation: Dissolved the powder in 10 ml Milli Q-H2O at 1mg/ml, and aliquoted to 1ml per 1.5 ml eppendorf tube, stored at -20oC. From this stock solution, prepare working solution. DAPI staining work solution (stored at 4oC): 2 µl (2 µg) stock solution in 1 ml (or adjusted according to your standard) staining buffer as described below: DAPI staining buffer (stored 4oC):

100 mM Tris, pH 7.4, 150 mM NaCl, 1 mM CaCl2•2H2O, 0.5 mM MgCl2, 0.1% Triton X-100. Sample preparation: Microspore released from anther or pellet from microspore culture by spinning down. Add sufficient amount of fresh prepared staining work solution, such as 200 µl per 1.5 ml eppendorf tube, pipette 10-15 µl staining work solution on slide, add glass cover, check under the UV microscopy for DAPI florescence.