Charcoal treatment of seawater to remove organics ( esp . vitamins ) .
Collect seawater for medium , filter through a 0.22 μm filter to remove cells and particles .
Autoclave or microwave the filtered water to sterilze .
Allow water to cool before treating with charcoal .
Pour the sterilzed water into a designated flask with stir bar that can contain at least 1L of liquid .
Wash activated charcoal .
Add 2.5g NaCl to 500mL of Milli Q water into a designated clean flask for charcoal washing .
The charcoal sticks so it is best to dedicate a flask for washing ( and other treatment steps ) .
Add 10g of activated charcoal to the NaCl2 solution .
Add stir bar .
Stir for at least 10min .
Recover the charcoal by filtration upon a glass fiber ( GF / F ) filter using a designated filter rig .
Use a scoopula to add washed charcoal to the sterilized seawater made in Step1 .
Take care not to rip the GF / F filter .
Obtain the remaining charcoal in the rig by swirling sterilzed water in the filtration rig and pouring that out into your flask with the sterilzed seawater ( and now washed charcoal ) .
Stir the washed charcoal plus sterilzed seawater for at least 30min , using a stir plate .
Remove charcoal from the seawater by filtering the mixture using a disposable filtration rig with a 0.2 μm filter .
After removing the charcoal and any larger than 0.2 μm particles , work in a sterile environment ( laminar flow hood recommended ) and aseptically add desired nutrient requirements to your seawater to complete your medium .
