Agar stab
In Microbiology, an agar stab is a tube containing bacteria immersed in a solid piece of agar, and is used for transport and storage. This method of Microbiological culture can be used to store bacteria for up to one year when kept in the dark at 4°C[1].
Preparing an agar stab
Preparing an agar stab is similar to preparing an agar plate for Microbiological culture. Sterile technique should be used in all steps to prevent contamination. The temperature and bacterial medium selected for your agar stab will vary depending on the identity of the bacterium. While an Agar plate typically uses 1.0 to 1.5% agar, an agar stab typically uses 0.6 to 0.7% agar[2][3].
To prepare an agar stab, solid bacterial medium is sterilized, transferred to a cryovial or screw top tube, and cooled to room temperature. The tube should be filled to 50% capacity. A single bacterial culture is picked from an Agar plate using an Inoculation loop (or a sterile toothpick, a sterile pipet tip, etc.) and plunged into the agar stab a few times. To ensure that the bacteria in the agar stab is viable, the cap is placed loosely on the tube to facilitate aeration and the agar stab is incubated at 37°C for 8 to 12 hours. After incubation, bacterial growth should be visible where the inoculation loop was stabbed into the agar. The stab can be stored at 4°C in the dark.
Reviving an agar stab
When a new agar stab is received, a sterile Inoculation loop is used to pick up some of the agar where the bacteria is visible as a streak. Next that agar is streaked onto an Agar plate and incubated overnight at 37°C (or a temperature appropriate for the bacterial species). The next day, a single colony is used to inoculate a liquid culture. The identity of the bacteria should be confirmed every time an agar stab is used.
Other methods of bacterial storage
Bacteria can also be stored as a Glycerol stock for long-term storage[4].
References
- ↑ Green, Michael R.; Sambrook, Joseph (February 2018). "Caring for Escherichia coli". Cold Spring Harbor Protocols. 2018 (2): pdb.prot101337. doi:10.1101/pdb.prot101337. ISSN 1940-3402.
- ↑ Elbing, Karen L.; Brent, Roger (January 2019). "Recipes and Tools for Culture of Escherichia coli". Current Protocols in Molecular Biology. 125 (1): e83. doi:10.1002/cpmb.83. PMC 6819147 Check
|pmc=value (help). PMID 30412361. - ↑ "Addgene: Pouring LB Agar Plates". www.addgene.org. Retrieved 2020-03-19.
- ↑ "Addgene: Protocol - How to Create a Bacterial Glycerol Stock". www.addgene.org. Retrieved 2020-03-19.
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