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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