Mimulus:Seed Sterilization

Seed Sterilization

Supplies

• 70% Ethanol
• 20% Bleach
• Sterile Water
• Seeds

Methods

1. Put seeds in an Eppendorf tube
2. Add 200uL of 70% EtOH to tube
3. Immediately add 600 uL of Water. Do only 2 tubes at a time, because the alcohol treatment is harsh and might affect germination if is left on seeds too long
4. Immediately suck off 600 uL of supernatant
5. Repeat steps 3 and 4 four times with water, not ethanol, to wash
6. Set aside tubes and repeat steps 1 to 5 for all the remaining tubes
7. Add 200 uL of 20% bleach to each tube
8. Immediatly remove bleach solution
9. Add 600 uL of water
10. Suck off 600 uL of the supernatant
11. Repeat steps 9 and 10 four or five times to wash