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