Efficient DNA Extraction from Nail Clippings Using the Protease Solution from Cucumis melo |
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Authors: | Shumi Yoshida-Yamamoto Sayaka Nishimura Teruko Okuno Miki Rakuman Yukio Takii |
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Affiliation: | (1) Department of Food Science and Nutrition, Mukogawa Women’s University, Nishinomiya Hyogo, 663-8558, Japan |
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Abstract: | Owing to the increasing importance of genomic information, obtaining genomic DNA easily from biological specimens has become
more and more important. This article proposes an efficient method for obtaining genomic DNA from nail clippings. Nail clippings
can be easily obtained, are thermostable and easy to transport, and have low infectivity. The drawback of their use, however,
has been the difficulty of extracting genomic material from them. We have overcome this obstacle using the protease solution
obtained from Cucumis melo. The keratinolytic activity of the protease solution was 1.78-fold higher than that of proteinase K, which is commonly used
to degrade keratin. With the protease solution, three times more DNA was extracted than when proteinase K was used. In order
to verify the integrity of the extracted DNA, genotype analysis on 170 subjects was performed by both PCR–RFLP and Real Time
PCR. The results of the genotyping showed that the extracted DNA was suitable for genotyping analysis. In conclusion, we have
developed an efficient extraction method for using nail clippings as a genome source and a research tool in molecular epidemiology,
medical diagnostics, and forensic science. |
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