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KMID : 1160220140420040311
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Mycobiology
2014 Volume.42 No. 4 p.311 ~ p.316
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An Easy, Rapid, and Cost-Effective Method for DNA Extraction from Various Lichen Taxa and Specimens Suitable for Analysis of Fungal and Algal Strains
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Park Sook-Young
Jang Seol-Hwa
Oh Soon-Ok
Kim Jung-A.
Hur Jae-Seoun
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Abstract
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Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive speciesidentification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacyin discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventionalmethods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cellwalls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. Thismethod involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, andprecipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from thefungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa weretested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearlydemonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.
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KEYWORD
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Lichens, Fungi, Algae, Genomic DNA, rRNA, Sequencing
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