GUS Staining of Pollen
(Janine’s protocol)

1. Add 200µl of acetone, vortex briefly.

2. Remove acetone and pipette into flat-bottom 96-well plate (polystyrene)

3. Centrifuge plate at 3000 x g for 3-5 minutes.

4. Invert plate carefully and blot dry with paper towels

5. Add 40µl of X-Gluc stain solution to each well (multi-pipettor) and incubate at 37°C overnight.

X-Gluc Solution

1 20 mg/ml X-Gluc concentration.  e.g. For a 1.5 ml tube (microfuge tube) dissolve 30 mg (.03 g) of X-Gluc in 1.5 ml DMSO (dimethyl sulfoxide).
2 Cover with foil.
3 Vortex well.
4 Keep at -20°C.

X-Gluc stain solution
*Note: 4ml of stain solution are necessary to stain 1 96-well plate.


 
Stock solution (10x)

50 mM ferrocyanide
50 mM ferricyanide

500mM NaPO4, pH 7

20mg/ml X-Gluc (in DMSO)

ddH2O

 Working solution (1ml)
 

100µl

100µl

25µl

775µl

1000µl = 1 ml
  • Cover with foil
  •  Keep at 4°C.