Processing of Microbial Isolates

Acceptable Samples
  • Solid media for bacterial and yeast species, frequently completed culture plates from the microbiology lab.
  • Collection of anaerobic, BL-3, fungal and virology isolates must be discussed with the specimen bank director regarding best means of collection.
Standard Procedure
  • Collect plates after completion of the culture.
  • Verify that the isolate is in pure culture, else set up new sheep’s blood agar plate with the strain and incubate at 37*C in 5% CO2 atmosphere or per appropriate conditions.
  • Once a pure culture has been verified, create duplicate frozen stocks of the isolate in cryovials with Tryptose Soy Broth media (TSB) and microbeads for adsorption of bacterial cells.
  • Use a swab to obtain a heavy inoculum of pure material from the plate and suspend in each vial.
  • Invert vials 5 times to mix and coat beads with bacteria.
  • For non-fastidious organisms, pipette off excess broth and discard.
  • For fastidious organisms leave excess TSB in the vials.
  • Store vials at -80*C.
Variations (added per request to study protocol).
  • Isolates can be frozen under other conditions, including with 20-50% glycerol in the media, or in media other than TSB.
  • Other containers: standard cryovials, 2″ crovials.
  • Additional aliquots (>2).
  • Agar slants can be made on chocolate agar or other media as needed, for pickup or shipment of isolates.
  • Special labeling of the tube (microbial species, sample source, etc.) as allowed in the investigator’s IRB protocol.