Arabidopsis EMS mutagenesis of seeds

 

Weight 0.2 gram seeds (10,000 seeds total) (20 microgram/seed).

Wash in 0.1% Tween 15'

Put into 15 ml ddH2O

Add 15-45 microliter (0.1% to 0.3%) EMS

Mix and incubate for 8-12 hrs (Rotating) in hood.

Remove the EMS (put EMS sol'n into 0.5M NaOH O/N, then dispose as regular waste)

Rinse the seeds once and then rinse them in 10 ml ddH2O 2-4 hrs.

Put all seeds into 100 ml 0.1% Agar (Autoclave agar, cool on ice, and let solidify for several hours first)

Sow the seeds using a P1000 pipetteman (1 ml per pot).