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

1. Add 300ml deionized, sterile H₂O to a 1 L beaker.
2. Add 83.72 g MOPS <-order more of this
3. Add 7.17 g Tricine
4. 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.
5. Add 0.14 g FeSO₄.7H₂O to 50 ml of deionized, sterile H₂O
6. Add 10 ml of the FeSO₄.7H₂O to the MOPS/Tricine solution.
7. Add 50 ml 1.9 M NH₄Cl
8. Add 10 ml 0.276 M K₂SO₄
9. Add 0.25 ml 0.02 M CaCl₂.2H₂O
10. Add 2.1 ml 2.5 M MgCl₂
11. Add 100 ml 5 M NaCl
12. Add 0.2 ml Micronutrient stock
13. Add 387 ml H₂O
14. 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.
15. Aliquot into 100 or 200 ml bottles and freeze at -20C.