Seeds from Arabidopsis, rice, tomato, pepper, etc., should be washed before we plate in the media agar or liquid (MS media). For this occasion, I just want to share the steps how to sterilize Arabidopsis seeds.
The whole steps are as follow:
- Transfer seeds into e-tube using small spoon or blade.
- Add 1 ml of Ethanol 100% into e-tube.
- Vortex for 3 minutes. The vortex strength should be in medium level (not too strong not too weak, but enough to mix the seeds with ethanol solution).
- Spin down briefly and discard the ethanol by pipetting.
- Add 1 ml of Sodium Hyperchloride solution (sodium hyperchloride:water = 1:6) into e-tube.
- Vortex for 3 minutes with medium strength.
- Spind down and discard sodium hyperchloride solution.
- Add 1 ml of sterilized water, mixing (vortex) for 3 minutes, spin down briefly, and change again with another sterilized water. Repeat this steps for 6 to 7 times.
- After last washing, add 1 ml sterilized water and store in 4 degree celcius for 2-3 days before plating into media.