Julius B. Lucks/M9 Supplemented Minimal Media

This protocol is a derivative of Endy:M9_media/supplemented. Endy protocol items in black, our protocol items in red.

Protocol

For 1L of media

• 500ml 2xM9 salts
  • used 11.28g of M9 Minimal Salts in 500ml of MiliQ-filtered H₂O
• 30ml 10 mg/ml thiamine hydrochloride
  • used 300 mg thiamine hydrochroride
• 10ml 40% glycerol
  • used 8ml 50% glycerol
• 20ml 10% casamino acids
  • wasn't sure if this was by weight or not. Assumed 10% by weight, so used 2g casamino acids.
• 20ml 0.1M MgSO₄
  • used 0.493 g (FW 246.49 g/mol)
• 200μl 0.5M CaCl₂
  • used the same (made 1ml 0.5 M CaCl₂ with 74 mg (FW 147.01 g/mol))
• 419.8ml sterile deionized H₂O
  • we made the above solution and filled to 1L with MiliQ-filtered H₂O

We combined the above ingredients, and filtered through a milipore steritop filter into a clean auto-claved 1L bottle. Stored at 4°C