Seed Sterilization and Seedling Growth on Plates in the Model Grass Brachypodium distachyon

Materials and Reagents

1. Square Petri dishes (Thermo fisher scientific, catalog number: 11708573 )
2. Microcentrifuge tubes (1.5 ml)
3. 3M^TM Micropore^TM Surgical Tape Standard (3M^TM, catalog number: 1530-0 )
4. Pipette tips
5. Brachypodium seeds
6. 70% ethanol
7. 95-100% ethanol
8. Sterile water
9. Household bleach (“regular strength” with 5.25% sodium hypochlorite)
10. Triton X-100 (e.g., Carl Roth, catalog number: 3051.2 )
11. Agar (e.g., Phyto agar, Duchefa Biochemie, catalog number: P1003.1000 )
12. Potassium hydroxide (KOH) pellets (e.g., Sigma Aldrich, catalog number: 30603-1KG )
13. MS (Murashige & Skoog, basal, without vitamins (Duchefa Biochemie, catalog number: M0221 ))
14. Sterilization solution (see Recipes)
15. ½ MS Media (see Recipes)
16. 1 M KOH (see Recipes)
    

Equipment

1. Pipette
2. Rocker (e.g., Benchmark Scientific Roto-Bot^TM)
3. Flame sterilized forceps with blunt end
4. 4 °C room or fridge
5. Growth chamber (e.g., Percival or Conviron)
6. Ethanol burner
   

Procedure