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Medium Detail Information

Medium No.
Medium for Strain M52
MgSO4·7H2O 0.5 g
CaCl2·2H2O 0.1 g
NH4Cl 0.1 g
KH2PO4 0.1 g
KCl 0.1 g
NaNO3 0.85 g
Na2S2O3·5H2O 2.5 g
Trace element mixture* 2 mL
Se/W solution** 2 mL
Bicarbonate solution*** 15-30 mL
Vitamin solution**** 1 mL
Distilled water 965 mL
  pH 7.0-7.2  

Mix ingredients except for bicarbonate and vitamin solutions. Dispense the medium ino suitable culture vessels under a stream of N2/CO2 (80/20) gas, seal with butyl rubber stoppers, and autoclave at 121°C for 20 min. Aspectically and anaerobically add bicarbonate and vitamin solutions. 

*Trace element solution SL-10

HCl (35%) 7 mL
FeCl3·6H2O 1.5 g
ZnCl2 70 mg
MnCl2·4H2O 100 mg
H3BO3 30 mg
CoCl2·6H2O 190 mg
CuCl2·2H2O 2 mg
NiCl2·2H2O 24 mg
Na2MoO4·2H2O 36 mg
Distilled water 1 L

First dissolve FeCl3 in concentrated HCl, dilute in water, and then add the other minerals.

**Se/W solution

Na2SeO4 4 mg
Na2WO4 4 mg
Distilled water 1 L

***Bicarbonate solution

8.4 g
Distilled water
100 mL

Saturated with CO2 gas. Tightly sealed i bottles filled with CO2 gas and then sterilized by autoclaving.

****Vitamin solution

Biotin 2 mg
Folic acid 2 mg
Pyridoxine-HCl (Vitamin B6) 10 mg
Thiamine-HCl·H2O (Vitamin B1) 5 mg
Riboflavin (Vitamin B2) 5 mg
Nicotinic acid 5 mg
D-Ca-Pantothenate 5 mg
Cyanocobalamin (Vitamin B12) 0.1 mg
p-Aminobenzoic acid 5 mg
Lipoic acid 5 mg
Distilled water 1 L

Adjust pH to 7.0 with NaOH.