Prerequisite Protocols

• MSA Plant Media

Materials

• MS Plant Media
• BAP 6-Benzylaminopurine 1mg/mL (150uL per 6 petri)
• NAA Napthaleneacetic acid 1mg/mL (15uL per 6 petri)
• Cefotaxime 100mg/mL (450uL per 6 petri)
• Kanamycin 100mg/mL (150uL per 6 petri)

Equipment

• Microwave Oven
• Micropipettes (200uL and 20uL)
• Laminar Flow Hood or Sterile Area (dead air box, bunsen sterile field, etc)
• Reusable Polypropylene Petri Dishes (PRE-STERILIZED)

Step By Step

1. Loosen the cap and melt down a 150mL aliquot of MS Plant Media in the microwave using a 1min on high followed by 30sec bursts, with stirring in between. Be careful to observer the process the first few times as each microwave is different. Be sure to use heat resistant gloves as the glass and media will be VERY HOT. Continue microwaving until no visible clumps of agar remain and the media spins freely. Look for trapped or slow rising bubbles as a sign of incomplete melting.
2. Cap the bottle and spray it with some 70% isopropanol and place the bottle under laminar flow hood or a sterile field.
3. Wait for the media to cool to about 55C (a hot cup of coffee you can just barely hold in your hands).
4. Dispense 150uL of BAP hormone into the solution.
5. Dispense 15uL of NAA hormone into the solution.
6. Dispense 450uL of Cefotaxime into the solution.