Team:UTK-Knoxville/media

M9 no Carbon

For 1L:

• 100mL 10X M9 Salts
• 1mL 1mg/mL thiamine HCl
• 50μL Trace Metals Solution
• 100μL 1M CaCl₂ (add last, while stirring)
• Filter Sterilize
• Glucose is added as appropriate from stock either into culture tubes or into media
• Add appropriate antibiotics to culture

10X M9 Salts

• 67.8g/L Na₂HPO₄
• 30g/L KH₂PO₄
• 5g/L NaCl
• 10g/L NH₄Cl
• Filter sterilize

M9 Plates

For 500mL:

• Autoclave 15g/L agar with ~448mL water
• Cool Down
• Add 50mL 10X M9 Salts
• 0.5mL 1M MgSO₄
• 0.5mL 1mg/mL thiamine HCl
• 25μL Trace Metals Solution
• 50μL 1M CaCl₂
• Add glucose from stock and appropriate antibiotics

M9 Medium+20% Sucrose

For 1L:

• Add 200g sucrose while heating and mixing with hotplate
• Turn off heat, continue stirring
• 4g/L glucose (to aid growth)
• 100mL 10X M9 Salts
• 1mL 1M MgSO₄
• 0.5mL 1mg/mL thiamine HCl
• 50μL Trace Metals Solution
• 100μL 1M CaCl₂
• Filter Sterilize
• Add appropriate antibiotics as necessary

LB

For 1L:

• Add 25g of LB broth and mix well with 1 liter H₂O
• Autoclave to sterilize