Have an old x-ray machine lying around. Use x-rays to effectively mutagenize spores for selection protocols
- X-ray machine
- Petri Dish, glass
- Plastic sandwich bag
- C-Fern spores
- Materials for spore sterilization and culture inoculation
1. Soak spores for 24 hours prior to sterilization and irradiation.
2. Sterilize using standard methods and place sterile spore suspension in a glass petri dish. Place dish inside plastic sandwich bag.
3. Irradiate spores with 25,000 Roentgens or about 555 Roentgens min-1 for 45 min. After irradiation sow spores as usual.
Under these conditions, spore germination at 10 days following sowing is about 80%; a screen with 10-5 M flurodeoxyuridine gives a mutant frequency of 2 X 10-5.