Glycerol & stock preparation
Updated 10/13/2023 09:47 AM
Introduction
Materials
- Material
- Falcon tubes of 50mL
- Glycerol 100%
- ddH2O
- syringe
- syring filter
- gun pipette
- serological pipettes
- equipment to achive sterile conditions
- For more details: Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual, Cabrera et al. 2020. Genes, Addgene
Procedure
Preparation of 10mL of glycerol solution 50% v/v
- Add 5mL of ddH20 inside a falcon tube of 50mL
- At the same falcon tube, add 5mL of glycerol 100%
- Mix well
- Take in the whole glycerol solution volume with a syringe
- Apply the syringe filter at the tip of the syringe
- Pour slowly the glycerol solution inside a new falcon tube of 50mL
Important Note:
1. the whole procedure mentioned above, should be conducted under sterile conditions using the fire of a gas burner.
2. check if the syringe fits the falcon tube and if its tip can reach the falcon tube bottom
Glycerol stock preparation
- For glycerol stocks, add the required volume of an overnight culture to a cryovial and the necessary glycerol volume to have a 25 % (v/v) final glycerol concentration. The final stock volume can be 1mL.
Glycerol stocks are important for long-term storage of strains and plasmids. Glycerol solution is essential for yeast and bacteria glycerol stocks preparation, aiming to enable long-term storage to -80°C.
Note: Glycerol solution concentration may change according to the cell type that needs to be stored.