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

被引：149

作者：

Izumitsu, Kosuke ^{[1
]}

Hatoh, Kanako ^{[1
]}

Sumita, Takuya ^{[1
]}

Kitade, Yuki ^{[1
]}

Morita, Atsushi ^{[1
]}

Gafur, Abdul ^{[2
]}

Ohta, Akira ^{[3
]}

Kawai, Masataka ^{[4
]}

Yamanaka, Takashi ^{[5
]}

Neda, Hitoshi ^{[5
]}

Ota, Yuko ^{[5
]}

Tanaka, Chihiro ^{[1
]}

机构：

[1] Kyoto Univ, Lab Environm Mycosci, Grad Sch Agr, Kyoto 6068502, Japan

[2] APRIL Fiber R&D, Pangkalan Kerinci 28300, Indonesia

[3] Shiga Forest Res Ctr, Yasu, Shiga 5202321, Japan

[4] Nara Forest Res Inst, Nara 6350133, Japan

[5] Forestry & Forest Prod Res Inst, Tsukuba, Ibaraki 3058687, Japan

来源：

MYCOSCIENCE | 2012年 / 53卷 / 05期

基金：

日本学术振兴会;

关键词：

Basidiomycetes; Colony PCR; Direct PCR; Fungi; ITS spacer region; FUNGI; GENE; TRANSFORMATION; SEQUENCES; TAXONOMY;

D O I：

10.1007/s10267-012-0182-3

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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.

引用

页码：396 / 401

页数：6

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