Tsubota Takahashi Nakahara Mohamed Deguchi 2005
Tsubota et al. (2005)
- Tsubota, H., Takahashi, K., Nakahara, M., Mohamed, H. & Deguchi, H. 2005. A simple procedure for DNA isolation using small quantities of lichen thallus. Lichenology 4: 25-28. [坪田博美・高橋奏恵・中原美保・H. Mohamed・出口博則: 少量の地衣体からのDNA簡易抽出法]
Abstract
Tsubota, H., Takahashi, K., Nakahara, M., Mohamed, H. & Deguchi, H. 2005. A simple procedure for DNA isolation using small quantities of lichen thallus. Lichenology 4: 25-28.
A simple protocol for DNA isolation from lichen thalli using small quantities (2 x 2 mm2 of samples is presented. The abundant polysaccharides and secondary metabolites present in a lichen thallus interfere with not only DNA isolation but also PCR amplification. The isolation procedure from the lichen thallus consists of the following short steps; (1) a fine-grind in a microcentrifuge tube; (2) extraction of ground tissue with the simple extraction buffer; (3) chloroform or CIA (chloroform and isoamylalcohol) extraction to remove impurities; and (4) adsorption of DNA with glass wool. DNA was successfully isolated from species of Sticta, Lobaria, Nephroma, and Pseudocyphellaria. DNA samples prepared by the present method were consistently amplifiable in LA-PCR as well as standard PCR reactions. This protocol also can be applicable for DNA isolation from other plant groups such as fungi, silamentous green algae and bryophytes.
- Key words
- lichens, fungi, green algae, bryophytes, DNA isolation, protocol, small quantities
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広島大学 / デジタル自然史博物館 / 植物 にもどる
