SSR-labeled fingerprint of shiitake mushroom Guangxiang strain, and application thereof
A technology of fingerprint and shiitake mushrooms, which is applied in the field of detection of shiitake mushroom strains, can solve problems such as affecting the enthusiasm for cultivation, affecting the rapid development of shiitake mushrooms, and the loss of mushroom farmers, and achieves the effects of good application prospects, short detection time and high accuracy.
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Embodiment 1
[0025] (1) Mycelia culture: transfer the shiitake mushroom strains to the potato glucose medium, culture in shake flasks in the dark at 23°C~25°C, and collect the mycelia after 3d~4d;
[0026] (2) Genomic DNA extraction: use CTAB (cetyltrimethylammonium bromide) method to extract the genomic DNA of the above hyphae, detect the concentration and purity of the total genomic DNA by ultraviolet spectrophotometry, and adjust the concentration of the sample DNA to be consistent;
[0027] The CTAB method for extracting the genomic DNA from mycelium includes:
[0028] ① Grind the mycelium of shiitake mushrooms freeze-dried in liquid nitrogen into powder, add 2×CTAB extract preheated at 65°C, keep warm at 65°C for 45-60min, and shake gently occasionally to mix well;
[0029] ②Centrifuge at 12000rpm, 4°C for 20min, and take the supernatant;
[0030] ③ Add a mixture of phenol, chloroform and isoamyl alcohol with a volume ratio of 25:24:1 to the above supernatant, mix gently for 10 minut...
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