Table T14. Media used for culturing prokaryotes, Expedition 330.
Media | Component | Mass/Volume/ Concentration | Treatment after mixing components |
---|---|---|---|
Artificial seawater (ASW) | 18.2 MΩ·cm MilliQ Water | 1 L | |
NaCl | 27.5 g | ||
MgCl2·6H2O | 12.16 g | ||
KCl | 0.72 g | ||
NaHCO3 | 0.2 g | ||
Tris-HCl | 3.2 g | ||
CaCl2·2H2O | 1.4 g | ||
NH4Cl | 0.05 g | ||
K2HPO4·3H2O | 0.05 g | ||
Trace elements solution | 1 mL | Before adding, adjust pH to 7.3 and autoclave | |
Trace vitamins solution | 1 mL | ||
Trace elements solution | ATCC trace mineral supplement | ||
Trace vitamins solution | 18.2 MΩ·cm MilliQ water | 1 L | |
Calcium pantothenate | 10 mg | ||
Niacin | 10 mg | ||
p-aminobenzoic acid | 10 mg | ||
Thiamine | 10 mg | ||
Riboflavin | 10 mg | ||
Pyroxidine | 10 mg | ||
Cobalamin (Vitamin B12) | 10 mg | ||
Thioctin (a-lipoic) acid | 10 mg | ||
Folic acid | 10 mg | ||
Biotin | 10 mg | Filter sterilize (no autoclave); store in darkness at 4°C | |
Autotrophic sulfur-oxidizing bacteria (ASOB) | Start with ASW | ||
Na2S2O3 | 10 mM | ||
Na2S | 100 µm | Purge with N2 prior to inoculating | |
Heterotrophic sulfur-oxidizing bacteria (HSO) | Start with ASOB | ||
Dextrose | 10 mM | ||
Autotrophic sulfate-reducing bacteria (ASR) | Start with ASW | ||
Na2SO4 | 60 mM | ||
Na2S | 500 µM | Purge with N2 prior to inoculating | |
Heterotrophic sulfate-reducing bacteria (HSR) | Start with ASR | ||
Lactate | 10 mM | ||
Acetate | 10 mM | If adding acetate as acetic acid, readjust pH to 7.3 | |
Autotrophic methanogen (AM) | Start with ASW | ||
Sodium molybdate | 5 mM | Filter sterilized solution | |
Na2S | 100 µm | Purge with N2 prior to inoculating | |
Heterotrophic methanogen (HM) | Start with AM | ||
Methanol | 10 mM | ||
Autotrophic Fe(III)-reducing bacteria (AIR) | Start with ASW | ||
Poorly crystalline Fe oxides | Goethite (<250 µm) or ferrihydrite are best; purge with N2 prior to inoculating | ||
Heterotrophic Fe(III)-reducing bacteria (HIR) | Start with AIR | ||
Lactate | 10 mM | ||
Acetate | 10 mM | If adding acetate as acetic acid, readjust pH to 7.3 | |
Autotrophic Fe(II)-oxidizing bacteria | Start with ASW | ||
FeCl2 | 1 mM | Add semi-daily, with some O2; keep microaerophilic (with added resazurin, solution should remain pink) | |
Marine broth (selects for heterotrophs; use 50%, 10%, and 1% media solution, diluted with autoclaved 3% NaCl) |
Difco marine broth from Sigma (Sigma-Aldrich, St. Louis, MO) |
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