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==Materials== *Sterile distilled water *70% ethanol *50% bleach/1 drop [http://en.wikipedia.org/wiki/Tween_20 Tween] 20: prepare 40 ml in a Falcon tube, use bleach that is less than two months old ==Procedure== #Transfer a small scoop of seed to an eppendorf tube; soak seed for 30 minutes in distelled water #Spin down seed briefly in microfuge #Decant water, add 500 uL 70% EtOH, vortex, spin #Decant EtOH, rinse with sterile water, spin and decant #Add 0.8 mL 50% bleach, vortex for 8-10 min, spin and decant #Add 0.8 mL sterile water, vortex, spin, decant #Repeat Step 6 six times #Leave seed in 0.5 mL of 0.12% sterile agar; allow to stratify at 4 degrees for at least 2 days ==Notes== *Keep the ethanol step as brief as possible-prolonged exposure to EtOH decreases germination *Multiple water washes are to remove all traces of bleach which also affects germination *Do the procedure in a hood in the tissue culture room *Store washed seed for stratification in refrigerator vegetable drawer in the prep room

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