Feb 25, 2022
Version 2
Algal Media Recipe from Cáceres Lab V.2
This document is a draft, published without a DOI.
- Carla Cáceres1,
- Isabella Oleksy2
- 1University of Illinois at Urbana-Champaign;
- 2Cary Institute for Ecosystems Studies
Document Citation: Carla Cáceres, Isabella Oleksy 2022. Algal Media Recipe from Cáceres Lab. protocols.io https://protocols.io/view/algal-media-recipe-from-c-ceres-lab-b5myq47wVersion created by Kira Monell
License: This is an open access document 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
Created: February 25, 2022
Last Modified: February 25, 2022
Document Integer ID: 58776
Abstract
This is a protocol to make media to grow algae in. In the Duffy Lab, this media is used to grow Ankistrodesmus falcatus. This recipe comes from the Cáceres Lab.
Algal Media Recipe from Cáceres Lab
Written by C. Caceres 2005
Reformatted by Isabella Oleksy 2013
Algal Nutrients
Stock solutions can be made up in 250mL, 500mL or 1L bottles. Use table to determine correct amount of nutrient to be added to desired amount of water. Use MilliQ water.
MICRONUTRIENTS
| A | B | C | D | E | |
| Name | Symbol | Add to 250mL | Add to 500mL | Add to 1L | |
| (1) EDTA Disodium Salt | Na2EDTA | 1.86g | 3.72g | 7.44g | |
| (2) Boric acid | H3BO3 | 0.62g | 1.24g | 2.47g | |
| (3) Maganese chloride Cobalt chloride Sodium molybdate | MnCl2 4H2O CoCl2 6H2O Na2MoO4 2H2O | 0.346g 0.005g 0.003g | 0.693g 0.010g 0.055g | 1.385g 0.019g 0.010g | |
| (4) Cupric sulfate Zinc sulfate | CuSO4 5H2O ZnSO4 7H2O | 0.0000465g** 0.23g | 0.000093g** 0.46g | 0.000186g** 0.920g | |
| (5) Ferric chloride | FeCl3 6H2O | 0.27g | 0.54g | 1.081g |
MACRONUTRIENTS
| A | B | C | D | E | |
| Name | Symbol | Add to 250mL | Add to 500mL | Add to 1L | |
| (6) Potassium phosphate | K2HPO4 | 4.35g | 8.71g | 17.42g | |
| (7) Sodium nitrate | NaNO3 | 42g | 85g | 170g | |
| (8) Calcium chloride | CaCl2 2H2O | 7.35g | 14.7g | 29.41g | |
| (9) Magnesium chloride | MgCl2 6H2O | 10.16g | 20.33g | 40.67g | |
| (10) Magnesium sulfate | MgSO4 7H2O | 12.33g | 24.65g | 49.3g |
**Because of the small amount needed, cupric sulfate should be made up at a 10x concentration and then diluted. Make a 0.0186g/L cupric sulfate solution and add 10mL to #4.
Store solutions at 4-6°C in polyethylene bottles.
A fresh batch of EDTA should be made every 6 months. Other solutions should be made fresh each year. When new solutions are made, containers should be scrubbed, rinsed, acid washed and rinsed well with DI water.
Algal media is made in either 5L or 2L bottles using 10 stock solutions and MilliQ water. The stock solutions are stored in the refrigerator and made as needed. Stock solutions should be remade every 1 year, with the exception of EDTA which should be remade every 6 months.
1)Fill bottles to ‘shoulder’ with fresh MilliQ water.
2)Using a clean Eppendorf repeater pipette for the stock solutions. Never put the Eppendorf pipette into the bottle of stock solution. Instead, pour the appropriate amount of stock solution into a clean beaker. Combine 1 mL/L of stock solution #1-5 IN NUMERICAL ORDER into a small beaker. Each bottle of medium you are making gets its own beaker. For each of the 5L bottles, you will use 5mL of each stock solution and for each 2L bottle you will use 2mL of each stock solution. Swirl well and let sit for 1 minute. Add each beaker to its own media bottle. There is no need to rinse the pipette between stock solutions.
3)Use the Eppendorf pipette to add 1 mL/L of stock solutions #6-10 directly to the media bottle (ie, 5mL of each stock solution for each 5L bottle and 2mL of stock solution for each 2L bottle).
4)Cover tightly with aluminum foil and place in an autoclavable container with about an inch of water.
5)Autoclave on the liquid setting for 30 minutes at 121°C.After autoclaving, remove and let cool. Check to be sure that precipitation of nutrients has not occurred.
6)When cool, attach to chemostat system.
7)Vitamins should not be autoclaved. Add separately each time new media is added. Use 1mL/L.