APPENDIX 7 - SILVER STAINING PROCEDURE

From Neilan et al, 1994

For 1 mm thick polyacrylamide gels

1. Wash gel twice with Milli-Q

2. Add 250 mL 10% ethanol (diluted in Milli-Q) for 5 minutes. Pour out.

3. Add 250 mL 1% nitric acid, keep on shaker for 3 minutes

4. Rinse with Milli-Q

5. Add 150 mL 0.2% AgNO3 and leave shaking for 20 minutes (or overnight)

6. Pour silver solution back into its container - it may be reused

7. Rinse gel and tray well with Milli-Q

8. Add developer:

Add ~100 mL; shake in indirect light until developer goes yellow in colour. Discard. Add the rest of the developer and shake until developed to desired extent.

9. Pour off and add 250 mL of 3% acetic acid. Leave for 5 minutes

10. Pour out acetic acid and rinse with 10% ethanol for 2 minutes

11. Pour out ethanol and store in 10% ethanol/ 7% glycerol solution