Note: Different groups of organisms sporulate better on different types of media (See Table A below) Remove a fresh slant of solid medium (SA, PD etc.) from the 4☌ refrigerator, allow to warm to room temperature and place in the BSC. Note: The method described below in section 11.1 can also be used for yeast isolates.įreezing Mold Isolates with heavy sporulation 11.1 Freezing Mold Isolates with heavy sporulation Step Using a sterile transfer pipette, transfer approximately 1.5 ml of the yeast suspension to each cryogenic vial. Make a heavy suspension of the yeast growth using a sterile inoculating loop or cotton swab by emulsifying the yeast growth (approximately 5 colonies) into the 15 mL graduated test tube. Using aseptic technique, pipette 9 mL of sterile 20% Glycerol solution to a sterile15 mL graduated conical tube. Handwriting should be legible and ink robust to tolerate -80☌. Labels should be sufficiently robust to tolerate temperature of -80☌ without disintegrating. Prepare screw capped cryogenic vials labelled with the date and isolate identification name/number. Store vials at -70☌ or as appropriate for your laboratory.Īlternate Method: Freezing Yeast Isolates 10.2 Alternate Method: Freezing Yeast Isolates Stepįollow Steps 1 through 4 in freezing method 10.1. Make a heavy suspension of the yeast growth using a sterile inoculating loop or cotton swab by emulsifying the yeast growth into each vial. Handwriting should be legible and ink robust to tolerate -80☌.Īdd 1.5 ml of sterile 20% glycerol to each vial. Printed labels should be sufficiently robust to tolerate temperature of -80☌ without disintegrating. The following steps should be performed in a BSC to ensure sterility. You should only see yeast cells under 40x.
#Penecillium aspergillus spore microscope free
Adjust incubation time as needed until 2-3mm colonies are seen.Ĭheck the isolate microscopically to verify that it is free of bacterial contamination. Culture must be in mid- to late- log phase growth.
Incubate the inoculated plate for 48 hours at 30☌. Streak the SCG plate or slant by taking a light inoculum from the yeast culture to be stored and inoculate a fresh plate.
Always allow the plate or slant to warm to room temperature.
Remove a fresh SCG agar plate or slant from the 4☌ refrigerator and place on the benchtop.
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