Creating Bacterial Glycerol Stocks for Long-term Storage
Bonnie Evans
Abstract
Bacterial glycerol stocks are important for long-term storage. Bacteria on LB agar plate can be stored at 4°C for a few weeks. However, if you want to store bacteria for a longer time, you will need to establish glycerol stocks. The addition of glycerol stabilizes the frozen bacteria, preventing damage to the cell membranes and keeping the cells alive. A glycerol stock of bacteria can be stored stably at -80°C for many years.
Before start
Make the 50% glycerol solution by diluting 100% glycerol in dH20 and autoclaving.
Steps
Steps
Grow an overnight culture of the selected strain
Inoculating a Liquid Bacterial Culture
Add 500 μL of the overnight culture to 500 μL of 50% glycerol in a 2 mL screw top tube or cryotube and gently mix
Shake the glycerol 5-6 times before freezing.
Freeze the glycerol stock tube at -80°C.
To recover bacteria from your glycerol stock, open the tube and use a sterile loop or pipette tip to scrape some of the frozen bacteria off of the top. Do not let the glycerol stock thaw!
Streaking and Isolating Bacteria on a LB Agar Plate
