Prerequisite Protocols

• MSA Solid Media

Materials

• BAP 6-Benzylaminopurine 1mg/mL (150uL per 6 petri)
• NAA Napthaleneacetic acid 1mg/mL (15uL 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 MSA Solid 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. Dispense 150uL of BAP hormone into the solution.
4. Dispense 15uL of NAA hormone into the solution.
5. Cap the bottle and stir contents manually to mix. Invert a few times. Use hot gloves as media is still molten.
6. Distribute molten media into petri dishes, ~25mL per dish. Recipe makes 6 thick dishes or 12 thin dishes.
7. Allow media to solidify under sterile field (~15mins). Cap, label with either acronym per petri or side stripe using marker according to your lab’s labeling system. Wrap in plastic wrap and store inverted at 4C in a refrigerator.

Notes

Lasts several months if stored at 4C. Replate every 2 weeks when in active use.