NMR/Minimal media

Trace Elements

In 800 ml dH₂O dissolve 5 g Na₂EDTA and correct to pH 7
Add the following in order, correcting to pH 7 after each:
- FeCl₃ (.6H₂0) 0.5 g (0.83 g)
- ZnCl₂ 0.05 g
- CuCl₂ 0.01 g
- CoCl₂.6H₂O 0.01 g
- H₃BO₃ 0.01 g
- MnCl₂.6H₂O (.4H₂0) 1.6 g (1.35 g)
Make up to 1 litre, autoclave and store at 4°C.

M9

Obtain M9 (or M9- for ¹⁵N labelling) from kitchen

Per litre, kitchen adds

6 g Na₂HPO₄
3 g KH₂PO₄
0.5 g NaCl

Then add

1 ml 1 M MgSO₄
200 µl 1 M CaCl₂
1 ml Thiamine (40 mg ml^-1 stock)
10 ml Trace Elements

Also add, as necessary

15 ml (¹³C-)Glucose (20 % Stock) (gives 0.3 % final)
1 g (¹⁵)NH₄Cl (if using M9-)

Tips

Grow from fresh transformation into rich media overnight, and use that to sub-culture labelling media at 1:50 for induction.