Knight:NuPAGE electrophoresis/Hybrid staining

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Overview

A fast protocol for visualizing bands on a polyacrylamide gel via coomassie-like staining.

Materials

Procedure

  1. Add 100mL deionized water in staining tray.
  2. Microwave staining tray loosely covered for 30 secs on high.
  3. Shake staining tray for 1 min on orbital shaker at room temperature.
  4. Discard water.
  5. Repeat steps 1-4 two more times.
  6. Add 20mL SimplyBlue SafeStain (enough to just cover the gel).
  7. Microwave staining tray loosely covered for 20 secs on high.
  8. Shake staining tray for 5 mins on orbital shaker at room temperature.
    • You should now be able to see most of the bands on the gel.
  9. Place staining tray with gel inside a ziploc bag to limit evaporation.
  10. Incubate overnight on orbital shaker at room temperature.
    • Overnight incubation will yield better sensitivity due to darker staining.
  11. Discard SimplyBlue SafeStain.
  12. Add 100mL deionized water.
  13. Add 2 kimwipes.
  14. Shake staining tray for 2-3 hours on orbital shaker at room temperature.
  15. Take gel picture.
  16. Add 20mL 20% NaCl for at least 5 mins.
    • Only do this step if you aren't planning on drying the gel! Otherwise, when the gel dries, you will see a white salt precipitate.
  17. Gel can be stored in salt solution for several weeks.
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