Protocol Citation: Maggi Mars 2025. L1 Media. protocols.io https://protocols.io/view/l1-media-dw3f7gjn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 02, 2025
Last Modified: January 02, 2025
Protocol Integer ID: 117575
Abstract
L1 media
Prep Bottles
Prep Bottles
Rinse 3x with DI water (Left Sink, with tubing) by filling app. 1/4 and swirling aggressively.
Rinse 3x with MilliQ water (dispenser, to the right of right sink) byt filling app. 1/4 and swirling aggressively.
Fill bottle with MilliQ to 1L line
Put autoclave tape on top of bottle, label MQ for milliQ water
loosen caps and put milliQ filled bottles into the solid autoclave bucket (not perforated), put bucket in autoclave and autoclave on liquid setting
Prepare Artificial Seawater
Prepare Artificial Seawater
empty a bottle of autoclaved MilliQ water
Add as much Instant Ocean to bottle as needed for the salinity you are making (40g for 36PSU)
Fill to one liter line with milliQ water
Put autoclave tape on top of bottle and label with the PSU being made
add a magnetic stirrer bar to bottle - shake aggressively to break up clumps. Put bottle on magnetic hot plate - turn on heat and stir until mostly all particulates dissolve
repeat for 4-5 bottles
loosen caps and put ASW filled bottles into the solid autoclave bucket (not perforated), put bucket in autoclave and autoclave on liquid setting
Let ASW cool enough to be handled comfortably
Shake to resuspend any particulates and/or put on magnetic stirrer until particulates are mostly dissolved
Empty out an autoclaved MilliQ bottle and attach a 0.2µm bottle-top filter (from shelves up and to the right from the sink)
Filter Seawater through the 0.2 µm filter using the vacuum pump next to the magnetic stirrer
Repeat for 4-5 bottles of Seawater (try to always do several bottles to get the most use out of the filter cups since they are big chunks of single-use plastic).
Make L1 Media
Make L1 Media
Add L1 media ingredients to the filtered, autoclaved ASW (N, P, Trace metals, vitamins, Silica)
Empty out an autoclaved MilliQ bottle and attach a 0.2µm bottle-top filter (from shelves up and to the right from the sink).
filter L1 media through 0.2 filter into the clean bottle.
Label filtered L1 media and store it in the fridge.
Repeat for 4-5 bottles of L1 media (try to always do several bottles to get the most use out of the filter cups since they are big chunks of single-use plastic).
Clean glassware
Clean glassware
Rinse and autoclave media bottles along the way, this protocol goes through a lot of autoclaved bottles ....