Endy:Neidhardt EZ Rich Defined
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.