Journal
MYCOSCIENCE
Volume 53, Issue 5, Pages 396-401Publisher
SPRINGER JAPAN KK
DOI: 10.1007/s10267-012-0182-3
Keywords
Basidiomycetes; Colony PCR; Direct PCR; Fungi; ITS spacer region
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Funding
- Research and Development Projects for Application in Promoting New Policies in Agriculture, Forestry, and Fisheries [21080]
- Japan Society for the Promotion of Science [22880017, 22380084]
- Grants-in-Aid for Scientific Research [22380084, 11J09315, 22880017] Funding Source: KAKEN
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We have optimized a simple and rapid preparation procedure for mushroom DNA extraction from colonies on media or from fruiting bodies for PCR amplification. The protocol combines microwaving twice for 1 min, cooling for 10 min, and centrifuging for 5 min. By using this procedure, more than 100 samples of mushroom DNA can be prepared within 1 h. The DNA obtained can be used for (1) identifying mushroom species by PCR and subsequent sequencing, (2) amplifying low copy number genes (at least 2,000 bp), and (3) screening genetic transformants. This technique will contribute to the mycology of mushroom species.
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