•  Fixative Solution: 50% methanol, 10% Acetic acid, 0.25% Glycerol
•  Solution A: 2% silver nitrate and 2% ammonium nitrate.
• Solution B: 10% tungstosilic acid
• Solution C: 3% formaldehyde
• Developer solution: 5% sodium carbonate
• Stopping reagent: 5% acetic acid.

 Procedure

1. Fix the Gel in the fixative solution for 20 minutes
2. Wash the gel in deionised water 2 times for 10 minutes each.
3. Mix 1 ml each of solutions A, B, and C and 7 ml of deionised water  and just  before use add  10 ml of developer solution  stain the gel in above solution in a  zipper bag till desired contrast is obtained. it usually takes 10- 15 minutes depending on the room temperature
4. When desired contrast is obtained, drain the staining solution and quickly add the stop solution to stop the staining.

Precautions

1. Handle the gels with gloved hands only.
2. Use deionised water.
3. Treat the glassware with concentrated nitric acid before use, it is preferable to use polyethylene zipper bags for the full procedure.
4. This receipe is for one 0.75mm thick gel, increase the quantities as per the thickness of gel.
5. To destain the gel put it in 3% hydrogen peroxide
   

Note

This protocol is based on the method of Gotlieb and Chavko

(updated 9/21/2000)