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Efficient DNA Extraction from Nail Clippings Using the Protease Solution from Cucumis melo

机译:使用黄瓜甜瓜蛋白酶溶液从指甲剪中高效提取DNA

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摘要

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.
机译:由于基因组信息的重要性日益增加,容易地从生物学标本中获得基因组DNA变得越来越重要。本文提出了一种从指甲剪中获得基因组DNA的有效方法。指甲剪很容易获得,具有热稳定性,易于运输,并且传染性低。然而,使用它们的缺点是难以从它们中提取基因组材料。我们使用从黄瓜甜瓜获得的蛋白酶溶液克服了这一障碍。蛋白酶溶液的角蛋白水解活性比通常用于降解角蛋白的蛋白酶K高1.78倍。用蛋白酶溶液提取的DNA是使用蛋白酶K提取的DNA的三倍。为了验证提取的DNA的完整性,通过PCR-RFLP和实时PCR对170位受试者进行了基因型分析。基因分型结果表明,提取的DNA适合于基因分型分析。总之,我们已经开发出一种有效的提取方法,可将指甲钳用作基因组来源,并将其用作分子流行病学,医学诊断和法医学的研究工具。

著录项

  • 来源
    《Molecular Biotechnology》 |2010年第1期|p.41-48|共8页
  • 作者单位

    Department of Food Science and Nutrition, Mukogawa Women’s University, Nishinomiya, Hyogo, 663-8558, Japan;

    Department of Food Science and Nutrition, Mukogawa Women’s University, Nishinomiya, Hyogo, 663-8558, Japan;

    Department of Food Science and Nutrition, Mukogawa Women’s University, Nishinomiya, Hyogo, 663-8558, Japan;

    Department of Food Science and Nutrition, Mukogawa Women’s University, Nishinomiya, Hyogo, 663-8558, Japan;

    Department of Food Science and Nutrition, Mukogawa Women’s University, Nishinomiya, Hyogo, 663-8558, Japan;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Nail clippings; Polymorphism; DNA extraction method; Cucumis melo; Keratin degradation;

    机译:指甲剪;多态性;DNA提取方法;甜瓜;角蛋白降解;

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