Feb 25, 2022
Vitamins for Algal Chemostats
This document is a draft, published without a DOI.
- 1University of Michigan
Document Citation: Meghan Duffy 2022. Vitamins for Algal Chemostats . protocols.io https://protocols.io/view/vitamins-for-algal-chemostats-b5m2q48e
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: 58778
Abstract
This a protocol to make vitamins for algal chemostats that are used to feed Daphnia and Ceriodaphnia in the Duffy Lab.
VITAMINS – CONTENTS
| A | B | C | |
| VITAMIN | mg/2L | Sigma Cat# | |
| 1. Biotin | 10 | B4501 | |
| 2. Thiamine | 200 | T4625 | |
| 3. Pyridoxine | 200 | P9755 | |
| 4. Pyridoxamine | 6 | P9380 | |
| 5. Calcium pantothenate | 500 | P2250 | |
| 6. B 12 | 2 | V2876 | |
| 7. Nicotinic Acid | 100 | N4126 | |
| 8. Nicotinamide | 100 | N3376 | |
| 9. Folic Acid | 40 | F7876 | |
| 10. Riboflavin | 60 | R4500 | |
| 11. Inositol | 180 | I5125 |
Since algae do not supply all the requirements for zooplankton culture, a mixture of eleven vitamins is added to the algal culture while it is in exponential growth phase.
To make the mixture, compounds are weighed on the analytical balance and combined in 2L volumetric flask containing approximately 1800 ml of MilliQ water. Cover the flask with aluminum foil or a box and allow to mix on a stir plate until the vitamins have dissolved into a clear yellow color. Fill to 2L with MilliQ water. A minimum volume of 2L is used in order to have a weighable amount of pyridoxamine and B 12.
The solution is then dispensed into dated 6 oz. Whirl paks and frozen.
As needed, vitamins are thawed and poured into sterile, microcentrifuge tubes. The tubes are then refrozen and stored in the dark since vitamins are sensitive to light and heat.
The mixture can kept frozen for more than a year and at 4-6 C for several weeks.
1-2 ml of vitamin mixture is normally added for each liter of media solution. Vitamins are added to the algae by withdrawing the desired amount using a syringe and injecting it into the silicone tubing while adding media to the algal culture.