Freezing Bacterial Cells
Categorized: Microbiological Techniques
- Grow the strain of bacteria to be frozen, in 5 to 10 mls of appropriate media, until an OD600 = 0.6-0.8 is reached OR aseptically (using a sterile swab) scrape bacteria from a freshly grown (24 – 48 hr) plate and resuspend in 2 to 5 mls of a rich broth such as Mueller Hinton or LBNS.
- Using a sterile pipette, add dimethyl sulfoxide (DMSO) so the final concentration is 7% (i.e. 70ul DMSO per 1 ml of cell suspension). Note: DMSO is considered a sterile solution but can be filter sterilized using a 0.22 um nylon filter.
- Vortex.
- Aseptically pipet 1.0 to 1.5 mls of bacterial suspension (with DMSO) into each cryovial. Make sure the cryovial is labelled with a waterproof, cryogenic pen (black color lasts better in the freezer). Freeze at -70oC.
Method Categories
- Biochemical Assays (16)
- Microbiological Techniques (8)
- Host-Pathogen Interactions (3)
- Functional Genomics (1)
- Immunology Based Methods (8)
- Peptides (2)
- Antibiotics and Antimicrobial Peptides (6)
- Bacterial Genetics (22)
- Protein Purification and Gel Electrophoresis (18)
- Liposome Methods (6)
- Outer Membranes (26)