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 .
