Demonstration of Ascospores

Acetate Agar 2, Gorodkowa, and the V-8 media are described by Yarrow (1998), and the enriched yeast extract-malt extract medium was originally described by van der Walt and van Kerken (1961). The staining procedure for spore cultures using malachite green is from Yarrow (1998). 

Place the flask in boiling water to dissolve the ingredients. Once dissolved, add distilled water to dilute to volume (lOOOmL) and autoclave at 121°C/250°F for 15 min. 

Once removed from the autoclave, temper the media in a water bath at 45°C/113°F to 50°C/122°F. 

Prepare pour plates and allow agar to solidify. 

Using a flamed and cooled loop, transfer a young culture (incubation time of 24 to 48 h) to the medium and incubate at 25°C/77°F for 2 to 3 days prior to examination with a microscope. 

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