Artificial Seawater Based AMP1 Medium
Dissolve each salt completely before adding the next one. 
Dispense into 500 ml acid-washed polycarbonate bottles.  
Autoclave 30 min. pH NaH2PO4 to 7.5 using 1M NaOH. 
Filter sterilize each solution using 0.2 µm syringe filter into new, sterile 50ml centrifuge tubes or acid washed and sterile polycarbonate bottles. 
Store at 4°C pH HEPES to 7.5 using 1M NaOH.  
Filter sterilize each solution using 0.2 µm syringe filter into new, sterile 15 or 50 ml centrifuge tubes or acid washed and sterile polycarbonate bottles. 
Store at 4°C. 
Dispense 5 ml of NaHCO3 and 0.5 ml HEPES per 500 ml bottle of Turk's Island Salt Mix when preparing final medium. 
Weigh out 0.2175 g Na2EDTA·2H2O using dust free paper. 
Transfer to acid washed 50 ml volumetric flask filled with 40 ml MQ-water. 
Dissolve EDTA by inverting flask several times. 
Weigh out 0.16 g FeCl3·6H2O using dust free paper. 
Dissolve iron chloride into same volumetric flask by inverting several times. 
Individually add and dissolve 50 µl each of the ZnSO4, CoCl2, MnCl2, Na2MoO4, NaSeO3, and NiCl2 Primary Trace Metal Stocks. 
Adjust volume to 50 ml mark with MQ-water. 
Filter through a 0.2 µm syringe filter into sterile, acid washed container in laminar flow hood. 
Store sterile stock at 4°C. 
Dispense 5 µl Stock Trace Metal Mix to 500 ml bottle of Turk's Island Salt Mix when preparing final medium.
