Endy:Neidhardt EZ Rich Defined
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See the main Neidhardt EZ Rich Defined for full details on how we make this media.
The kitchen makes up the 10X MOPS mixture for us. Here is the protocol -
Making 1 L 10x MOPS mixture
- Add 300ml deionized, sterile H2O to a 1 L beaker.
- Add 83.72 g MOPS <-order more of this
- Add 7.17 g Tricine
- Add 10M KOH to a final pH of 7.4 (10 to 20 ml) <-make sure the kitchen has a stock of this. They don't seem to have it anymore. Also, they don't have a pH meter so they need to use the Sauer labs for this.
- Add 0.14 g FeSO4.7H2O to 50 ml of deionized, sterile H2O
- Add 10 ml of the FeSO4.7H2O to the MOPS/Tricine solution.
- Add 50 ml 1.9 M NH4Cl
- Add 10 ml 0.276 M K2SO4
- Add 0.25 ml 0.02 M CaCl2.2H2O
- Add 2.1 ml 2.5 M MgCl2
- Add 100 ml 5 M NaCl
- Add 0.2 ml Micronutrient stock
- Add 387 ml H2O
- Filter sterilize with a 0.2 micron filter. The kitchen doesn't have these, we need to bring one. They do have a vacuum line.
- Aliquot into 100 or 200 ml bottles and freeze at -20C.