1000x vitamin solution for yeast nitrogen base
The recipe described here is based on the description from the John McCusker lab. In contrast to the original recipe I have dissolved pyridoxine and thiamine in water instead of ethanol. Therefore I have added 4 ml EtOH to the ethanol-soluble vitamins in order to keep the volume the same. I have not checked if it is necessary to assure that all vitamins can be dissolved.
Volumes for the preparation of 50 ml of a 1000x vitamin stock solution.
|Compound||Stock solution||Vol. for 50 ml 1000x stock|
|biotin||0.005 g in 50 ml 95% EtOH||1 ml|
|calcium pantothenate||0.1 g in 10 ml H2O||2 ml|
|folic acid||0.0025 g in 50 ml H2O (warm)||2 ml|
|p-aminobenzoic acid||0.1 g in 10 ml 95% EtOH||1 ml|
|pyridoxine HCl||0.1 g in 10 ml H2O||2 ml|
|thiamine HCl||0.1 g in 10 ml H2O||2 ml|
|riboflavin 5' monophosphate||-||10 mg|
- remove the individual stock solutions from the refrigerator, may have to warm to get them back into solution
- combine all 95% EtOH vitamin solutions (2 ml), add 4 ml 95% EtOH
- Add 10 mg riboflavin 5' monophosphate (sodium salt dihydrate) to 95% EtOH mixture
- Combine the filtersterilized, water-dissolved vitamin solutions (11 ml) with 33 ml sterile H2O
- Combine the sterile H2O solution with the 95% EtoH solution/suspension for a final volume of 50 ml.