Seed sterilization
1. Create a solution of soap solution consisting of
a. 40 mL Clorox
b. 20 mL 95% ethanol
c. 40 mL water (from faucet)
d. 1 drop Triton
2. Put seeds into 15 mL Falcon test tubes and arrange tubes on a rack
a. It is best not to do more than 10 tubes at a time
3. Pour 2 – 6 mL of soap solution into tubes with a disposable pipette
a. The volume you add will depend on how many seeds you have. There should be
enough soap for the seeds to be adequately suspended while shaking.
4. Agitate constantly for 3 minutes so seeds are suspended in solution
5. Remove the soap solution either by:
a. Pouring off the solution very carefully to avoid losing seeds OR
b. Create a vacuum apparatus with items below and use a thin pipette to remove the
solution with minimal seed loss
i. Large Erlenmeyer flask
ii. Rubber cork with two holes
iii. Two lengths of rubber hose
iv. Thin glass Pasteur pipette
6. Wash 4 more times with sterilized, de-ionized water for 3 minutes each time
a. On the last wash, the water should no longer be soapy
7. Pour the sterilized seeds onto a sterile petri plate and let dry in a particle–free, horizontal
fume hood.
a. Since the seeds will stick to the sides of the test tube, it is best to use some form
of scooper. Be sure to sterilize in ethanol between uses and dry with a KimWipe