Rapid and simple preparation of mushroom DNA directly from colonies and fruiting bodies for PCR

Kosuke Izumitsu; Kanako Hatoh; Takuya Sumita; Yuki Kitade; Atsushi Morita; Chihiro Tanaka; Abdul Gafur; Akira Ohta; Masataka Kawai; Takashi Yamanaka; Hitoshi Neda; Yuko Ota

doi:10.1007/S10267-012-0182-3

Short communication

Rapid and simple preparation of mushroom DNA directly from colonies and fruiting bodies for PCR

Kosuke Izumitsu, Kanako Hatoh, Takuya Sumita, Yuki Kitade, Atsushi Morita, Chihiro Tanaka, Abdul Gafur, Akira Ohta, Masataka Kawai, Takashi Yamanaka, Hitoshi Neda, Yuko Ota

Author information

Kosuke Izumitsu
Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Kanako Hatoh
Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Takuya Sumita
Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Yuki Kitade
Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Atsushi Morita
Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Chihiro Tanaka
Laboratory of Environmental Mycoscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Abdul Gafur
APRIL Fiber R&D, Pangkalan Kerinci 28300, Indonesia
Akira Ohta
Shiga Forest Research Centre, Yasu, Shiga 520-2321, Japan
Masataka Kawai
Nara Forest Research Institute, Takatori, Nara 635-0133, Japan
Takashi Yamanaka
Forestry and Forest Products Research Institute, Tsukuba, Ibaraki 305-8687, Japan
Hitoshi Neda
Forestry and Forest Products Research Institute, Tsukuba, Ibaraki 305-8687, Japan
Yuko Ota
Forestry and Forest Products Research Institute, Tsukuba, Ibaraki 305-8687, Japan

Keywords: Basidiomycetes, Colony PCR, Direct PCR, Fungi, ITS spacer region

JOURNAL OPEN ACCESS

2012 Volume 53 Issue 5 Pages 396-401

DOI https://doi.org/10.1007/S10267-012-0182-3

Details

Abstract

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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