Medium data

Search for medium no. = [1100].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1100   CTM MEDIUM

CTM medium basis:
Solution 1 (see below)  20.0     ml
Solution 2 (see below)  3.0     ml
Solution 3 (see below)  2.0     ml
Solution 4 (see below)  2.5     ml
HEPES  2.4     g
Mix all ingredients in 1.0 L medium bottle. Add distilled water close to 1.0 L. Adjust pH to 7.0 with 2 M KOH and fill up to 1.0 L. Autoclave at 121C for 40 min. After autoclaving, seal medium bottle immediately in order to avoid oxygenation. Cool down to around 70C. Add 30 ml filter-sterilized Mixed solution (see below) to the autoclaved CTM medium basis for completing the CTM medium.
Solution 1 (50 x stock):
MgSO4·7H2O  3.75    g
CaCl2·2H2O  1.80    g
Citric acid  0.30    g
0.25 M EDTA·2Na solution (pH 8.0)  0.60    ml
Trace metal solution (see below)  50.0     ml
Distilled water  1.0     L
Trace metal solution:
H3BO3  2.86    g
MnCl2·4H2O  1.81    g
ZnSO4·7H2O  0.222   g
Na2MoO4·2H2O  0.39    g
CuSO4·5H2O  0.079   g
Co(NO3)2·6H2O  0.049   g
Distilled water  1.0     L
Solution 2:
K2HPO4  15.3     g
Distilled water  1.0     L
Solution 3:
Ferric ammonium citrate  12.0     g
Distilled water  1.0     L
Solution 4:
2-Oxoglutarate  14.6     g
Distilled water  1.0     L
Mixed solution:
NaHCO3  0.625   g
Sodium thioglycolate  0.125   g
Solution 5 (see below)  0.5     ml
Solution 6 (see below)  10.0     ml
Vitamin solution A (see below)  0.25    ml
Vitamin solution B (see below)  0.25    ml
Add distilled water to bring final volume to 30 ml.
Solution 5:
Bacto peptone (BD-Difco)  10.0     g
Distilled water  100.0     ml
Solution 6:
L-Alanine  100.0     mg
L-Arginine  100.0     mg
L-Asparagine  100.0     mg
L-Aspartic acid  100.0     mg
L-Cysteine  100.0     mg
L-Glutamic acid  100.0     mg
L-Glutamine  100.0     mg
Glycine  100.0     mg
L-Histidine  100.0     mg
L-Isoleucine  100.0     mg
L-Leucine  100.0     mg
L-Lysine  100.0     mg
L-Methionine  100.0     mg
L-Phenylalanine  100.0     mg
L-Proline  100.0     mg
L-Serine  100.0     mg
L-Threonine  100.0     mg
L-Tryptophan  100.0     mg
L-Tyrosine  100.0     mg
L-Valine  100.0     mg
Distilled water  400.0     ml
Vitamin solution A (1000 x stock):
Biotin  10.0     mg
Riboflavin  10.0     mg
Thiamine·HCl  100.0     mg
Thiamine pyrophosphate  100.0     mg
L-Ascorbic acid  100.0     mg
Calcium pantothenate  100.0     mg
Folic acid  100.0     mg
Nicotinamide  100.0     mg
Nicotinic acid  100.0     mg
p-Aminobenzoic acid  100.0     mg
Pyridoxine·HCl  100.0     mg
Lipoic acid  100.0     mg
β-NAD  100.0     mg
10 mM Potassium phosphate buffer (pH 7.2)  100.0     ml
Vitamin solution B (1000 x stock):
Vitamin B12  100.0     mg
Distilled water  100.0     ml