How to recover bacterial isolates from long term storage

This blog post contains general information regarding the recovery of a bacterial isolate from a frozen or lyophilized storage condition.

To recover the isolate follow the step wise procedure as mentioned below;

1. Remove the frozen cultures from the freezer and place them on dry ice or into an alcohol and dry-ice bath;
2.  Transfer to a laboratory safety cabinet or a clean area if a cabinet is not available.
3. Scrape the top-most portion of the culture using a sterile loop
4. Transfer to a growth medium without contaminating the top or inside of the vial.
5. Re-close the vial before the contents completely thaw, and return the vial to the freezer; with careful technique, transfers can be successfully made from the same vial several times.
6. Incubate 18–24 hours at 35–37°C; perform at least one subculture before using the isolate to inoculate a test.