Preparing the culture
Grow sporulating strains on solid media as for freeze-drying; grow
nonsporulating strains on either solid or liquid medium. If the
mycelium is easily broken, grow the culture on agar in test tubes,
scrape with a pipette, and suspend the fragments in sterile 10%
glycerol. Dispense 0.5 mL into each plastic vial. If the mycelium is
sticky, will not break up, or grows embedded in the agar, grow the
culture on agar in plates, cut out plugs containing new growth
(hyphal tips) with a sterile cork borer, and place three or four plugs
into each plastic freezing vial with approximately 0.4 mL of 10%
glycerol.
Freezing methods
To ensure long-term viability of fungal cultures, ATCC recommends
freezing and storage at liquid nitrogen temperatures. Storage in
the liquid itself is not always convenient or safe. Storage in liquid
nitrogen vapor is a more practical alternative. It is critical, however,
to constantly monitor the liquid level of the liquid nitrogen freezer
to ensure that material is maintained below –130°C. Storage at
warmer temperatures can compromise the stability of many strains.
Preparing the cultureGrow sporulating strains on solid media as for freeze-drying; grownonsporulating strains on either solid or liquid medium. If themycelium is easily broken, grow the culture on agar in test tubes,scrape with a pipette, and suspend the fragments in sterile 10%glycerol. Dispense 0.5 mL into each plastic vial. If the mycelium issticky, will not break up, or grows embedded in the agar, grow theculture on agar in plates, cut out plugs containing new growth(hyphal tips) with a sterile cork borer, and place three or four plugsinto each plastic freezing vial with approximately 0.4 mL of 10%glycerol.Freezing methodsTo ensure long-term viability of fungal cultures, ATCC recommendsfreezing and storage at liquid nitrogen temperatures. Storage inthe liquid itself is not always convenient or safe. Storage in liquidnitrogen vapor is a more practical alternative. It is critical, however,to constantly monitor the liquid level of the liquid nitrogen freezerto ensure that material is maintained below –130°C. Storage atwarmer temperatures can compromise the stability of many strains.
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