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DSMZ

JCM 1100 . CTM MEDIUM

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Equipment needed: Autoclave Filter
Final pH: 7.0
Select a strain (optional):

ml
Compound Amount Unit
Solution 1 (50 x stock) 20.0 ml
Solution 2 3.0 ml
Solution 3 2.0 ml
Solution 4 2.5 ml
HEPES 2.4 g
1 CTM medium basis:
2 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) #

1051
Compound Amount Unit
MgSO4 x 7 H2O 3.75 g
CaCl2 x 2 H2O 1.80 g
Citric acid 0.30 g
Na2-EDTA
(pH 8.0, 0.25 M) 		0.60 ml
Trace metal solution 50.00 ml
Distilled water 1000.00 ml

Trace metal solution #

1000
Compound Amount Unit
H3BO3 2.860 g
MnCl2 x 4 H2O 1.810 g
ZnSO4 x 7 H2O 0.222 g
Na2MoO4 x 2 H2O 0.390 g
CuSO4 x 5 H2O 0.079 g
Co(NO3)2 x 6 H2O 0.049 g
Distilled water 1000.000 ml

Solution 2 #

1000
Compound Amount Unit
K2HPO4 15.3 g
Distilled water 1000.0 ml

Solution 3 #

1000
Compound Amount Unit
Ferric ammonium citrate 12 g
Distilled water 1000 ml

Solution 4 #

1000
Compound Amount Unit
2-Oxoglutarate 14.6 g
Distilled water 1000.0 ml

Mixed solution #

11
Compound Amount Unit
NaHCO3 0.625 g
Sodium thioglycolate 0.125 g
Solution 5 0.500 ml
Solution 6 10.000 ml
Vitamin solution A (1000 x stock) 0.250 ml
Vitamin solution B (1000 x stock) 0.250 ml
1 Add distilled water to bring final volume to 30 ml.

Solution 5 #

100
Compound Amount Unit
Bacto peptone
(BD-Difco) 		10 g
Distilled water 100 ml

Solution 6 #

400
Compound Amount Unit
L-Alanine 100 mg
L-Arginine 100 mg
L-Asparagine 100 mg
L-Aspartic acid 100 mg
L-Cysteine 100 mg
L-Glutamic acid 100 mg
L-Glutamine 100 mg
Glycine 100 mg
L-Histidine 100 mg
L-Isoleucine 100 mg
L-Leucine 100 mg
L-Lysine 100 mg
L-Methionine 100 mg
L-Phenylalanine 100 mg
L-Proline 100 mg
L-Serine 100 mg
L-Threonine 100 mg
L-Tryptophan 100 mg
L-Tyrosine 100 mg
L-Valine 100 mg
Distilled water 400 ml

Vitamin solution A (1000 x stock) #

100
Compound Amount Unit
Biotin 10 mg
Riboflavin 10 mg
Thiamine HCl 100 mg
Thiamine pyrophosphate 100 mg
L-Ascorbic acid 100 mg
Calcium pantothenate 100 mg
Folic acid 100 mg
Nicotinamide 100 mg
Nicotinic acid 100 mg
p-Aminobenzoic acid 100 mg
Pyridoxine hydrochloride 100 mg
Lipoic acid 100 mg
beta-NAD 100 mg
Potassium phosphate buffer
(pH 7.2, 10 mM) 		100 ml

Vitamin solution B (1000 x stock) #

100
Compound Amount Unit
Vitamin B12 100 mg
Distilled water 100 ml