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

2012; Elsevier BV; Volume: 53; Issue: 5 Linguagem: Inglês

10.1007/s10267-012-0182-3

1618-2545

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

Plant Pathogens and Fungal Diseases

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.