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

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 1min, cooling for 10min, and centrifuging for 5min. By using this procedure, more than 100 samples...

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Veröffentlicht in:Mycoscience 2012-09, Vol.53 (5), p.396-401
Hauptverfasser: Izumitsu, Kosuke, Hatoh, Kanako, Sumita, Takuya, Kitade, Yuki, Morita, Atsushi, Tanaka, Chihiro, Gafur, Abdul, Ohta, Akira, Kawai, Masataka, Yamanaka, Takashi, Neda, Hitoshi, Ota, Yuko
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Sprache:eng
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Zusammenfassung: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 1min, cooling for 10min, and centrifuging for 5min. By using this procedure, more than 100 samples of mushroom DNA can be prepared within 1h. The DNA obtained can be used for (1) identifying mushroom species by PCR and subsequent sequencing, (2) amplifying low copy number genes (at least 2,000bp), and (3) screening genetic transformants. This technique will contribute to the mycology of mushroom species.
ISSN:1340-3540
1618-2545
DOI:10.1007/S10267-012-0182-3