Endy:M9 media/supplemented: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Barry Canton (talk | contribs) |
Barry Canton (talk | contribs) No edit summary |
||
Line 1: | Line 1: | ||
''This is our protocol for making [[M9 medium/supplemented|supplemented M9 medium]].'' | |||
==Component Concentrations== | ==Component Concentrations== | ||
*1X [[M9 salts]] | *1X [[M9 salts]] |
Revision as of 18:43, 13 September 2006
This is our protocol for making supplemented M9 medium.
Component Concentrations
- 1X M9 salts
- 1mM thiamine hydrochloride
- 0.4% glycerol
- 0.2% casamino acids
- 2mM MgSO4
- 0.1mM CaCl2
Protocol
For 1L of 1X media
- 500 mL 2X M9 minimal salts ∗
- 30 mL 10 mg/mL thiamine
- Dissolve 10 mg per mL of H2O
- Use a 0.22 μm filter to filter-sterilize
- Light-sensitive: store covered
- 10 mL 40% glycerol ∗
- 20 mL 10% Casamino acids ∗
- 20 mL 0.1M MgSO4 ∗
- 200 μL 0.5M CaCl2 ∗
- 419.8 mL H2O
Combine above solutions using sterile technique. (May notice some precipitation during preparation but precipitate should go back into solution once volume is brought up to 1L with sterile H2O.)
Add antibiotic as appropriate and store at 4°C
∗ Can be obtained in the media room