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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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