Team:UTK-Knoxville/media
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Revision as of 21:15, 15 July 2014
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