Sieve the dry seed through a fine sieve (we found a fine plastic or stainless steel tea strainer to be suitable) on to paper to remove all larger debris.
When sieving avoid drafts .
Take a small vial (5 to 20mLs) with a screw top.
Transfer seed into dry vial.
Add a few drops of 70% alcohol (or methylated spirits) to just wet the seed
Quickly add enough 3% peroxide to cover seed plus the height of seed layer above seed (eg 2mm seed – 4mm peroxide)
Soak inside of vial lid with peroxide to sterilize it and screw it on firmly.
Surface sterilise the exterior of the vial with alcohol solution or bleach solution.
Transfer to sterile work area
Shake for 10 minutes
Tip all contents of vial onto sterile agar (under aseptic conditions) – the mother flask should be 500 to 750mls in size – we use ¼ turn twist top jars with 2 layers of coffee filter paper cut to just perfectly fit in the lid. Sterile mother flask should contain about 120 to 150 mL of medium.
Leave flask in dark for 12 to 24hours then put under lights to germinate and grow.
|
