Have an old x-ray machine lying around. Use x-rays to effectively mutagenize spores for selection protocols

 

Materials

  • X-ray machine
  • Petri Dish, glass
  • Plastic sandwich bag
  • C-Fern spores
  • Materials for spore sterilization and culture inoculation¬†

Methods

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.