Purpose: Growth of bacteria at isotope labeled media to obtain proteins suitable for NMR analysis
Notes: when the iron stock starts to precipitate, make a fresh one (tube will turn brownish).
|
Materials |
1 litre medium |
100 ml medium |
|
Na2HPO4 |
6.0 gram |
0.6 gram |
|
KH2PO4 |
3.0 gram |
0.3 gram |
|
NaCl |
0.5 gram |
50 mg |
|
15NH4Cl |
2 gram |
200 mg |
|
MgSO4 (1M) |
2.0 ml |
0.2 ml |
|
CaCl2 (1M) |
10 ul |
1.0 ul |
|
FeSO4 (0.01M) |
1 ml |
100 ul |
|
13C-Glucose (when doubly labeled) |
2.0 gram |
0.2 gram |
|
12C-Glucose (when singly labeled) |
5.0 gram |
0.5 gram |
|
Thiamine (0.5 mg�ml-1) |
10 ml |
1.0 ml |
|
*Micronutrients (1000x) |
1.0 ml |
100 ul |
|
*Vitamin Supplements (1000x) |
1.0 ml |
100 ul |
|
*Chlorophenicols (1000x) |
1.0 ml |
100 ul |
|
Materials |
1000x stock concentration |
|
Ammonium molibdate |
3�10-6 M |
|
Boric acid |
4�10-4 M |
|
Cobalt chloride |
3�10-5 M |
|
Copper sulphate |
1�10-5 M |
|
Manganese chloride |
8�10-5 M |
|
Zinc chloride |
1�10-5 M |
|
Materials |
1 litre |
100 ml |
|
D-Biotin |
1.0 g |
100 mg |
|
Choline Chloride |
0.5 g |
50 mg |
|
Folic Acid |
0.5 g |
50 mg |
|
Myoinositol |
1.0 g |
100 mg |
|
Nicatinamide |
0.5 g |
50 mg |
|
Panthotenic Acid |
0.5 g |
50 mg |
|
Pyridoxal HCl |
0.5 g |
50 mg |
|
Riboflavin |
50 mg |
5 mg |
|
Thiamine HCl |
0.5 g |
50 mg |
Store in aliquots at –20� C and protect from light.