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KP7 buffer* 5ml
MgCl2·6H2O 3.05g
CaCl2·2H2O 0.15g
NH4Cl 0.54g
Na2SO4 0.3g
NaCl 30g
Bacto Yeast Extract (Difco) 0.02g
Sodium acetate 0.8g
Trace elements solution** 2ml
Vitamin solution*** 2ml
Resazurin 1mg
Na2CO3 1.5g
Na2S·9H2O 0.5g
Cysteine-HCl 0.5g
Distilled water 1L
pH unadjusted
Mix ingredients except KP7 buffer, vitamin solution, Na2CO3, Na2S·9H2O and cysteine-HCl. Dispense the medium into suitable culture vessels under a stream of H2/CO2 (80/20) and seal with butyl rubber stoppers. Separately autoclave KP7 buffer, Na2S·9H2O (5% solution) and cysteine-HCl (5% solution) under a N2 atmosphere. Sterilize vitamin and 10% Na2CO3 solutions by filtration. Prior to inoculation, aseptically and anaerobically add KP7 buffer, vitamin, Na2CO3 and Na2S·9H2O solutions. Pressurize the inoculated vessels to 150 kPa with H2/CO2.
KP7 buffer
KH2PO4 53g
K2HPO4 106g
Distilled water 1L
Autoclaved under N2.
**Trace elements solution
Nitrilotriacetic acid (NTA) 12.8g
FeCl3·6H2O 1.35g
MnCl2·4H2O 0.1g
CoCl2·6H2O 0.024g
CaCl2·2H2O 0.1g
ZnCl2 0.1g
CuCl2·2H2O 0.025g
H3BO3 0.01g
Na2MoO4·2H2O 0.024g
NaCl 1g
NiCl2·6H2O 0.12g
Na2SeO4 0.004g
Na2WO4 0.004g
KAl(SO4)2·12H2O 0.02g
Distilled water 1L
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0.
***Vitamin solution
Biotin 2mg
Folic acid 2mg
Pyridoxine-HCl 10mg
Thiamine-HCl 5mg
Riboflavin 5mg
Nicotinic acid 5mg
Ca-pantothenate 5mg
p-Aminobenzoic acid 1mg
Vitamin B12 0.01mg
Distilled water 1L

250g/瓶

常溫

3年