Melanie,
Here's my recipe for MJ media. It orginally came from a Montelione
paper, JBNMR 7 (1996) 131-141
Trace Elements Stock Solution (sterile filter):
FeCl3 * 6H2O
16.2g
ZnSO4 * 7H2O
2.4g
CoCl2 * 6H2O
4.2g
Na2MoO4 * 2H2O
4.2g
CuSO4 * 5H2O
4.8g
H3BO3
1.2g
MnSO4
3.0g
HCl (conc)
30 ml
H2O
570 ml
Vitamin Stock Solution (adjusted to pH 7.2, filter sterilized)
Pantothenic acid, calcium salt 0.4g
Choline chloride
0.4g
Folic Acid
0.4g
Myo-inositol
0.8g
Nicotinamide
0.4g
Pyridoxal hydrochloride
0.4g
Riboflavin
0.04g
Thiamine hydrochloride
0.4g
H2O
800ml
And to finally make the media:
1 L base media:
2.5g NH4Cl
9g KH2PO4
6g K2HPO4
0.5g sodium citrate
adjust to pH 6.6 and autoclave
After cooling, add a filter sterilized solution of the following:
3.0g glucose
1ml trace metal solution
0.65ml vitamin solution
0.07g thiamine
1g MgSO4
Requisite antibiotic for selection
In practice, we only made the trace metal solution and use Eagle Basal
Vitamin solution instead of the vitamin solution listed in the paper.
Before I tried this recipe I was using M9 with a different trace metal
solution and Eagle vitamin solution. My previous mix really wasn't very
different, but switching to this MJ media increased my isolated protein
yield from 0.5mg/l to 7mg/l.
Hope this helps!
Roger