Efficient DNA extraction from nail clippings using the protease solution from Cucumis melo

Mol Biotechnol. 2010 Sep;46(1):41-8. doi: 10.1007/s12033-010-9273-6.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Cucumis melo / enzymology*
  • DNA / isolation & purification*
  • Deoxyribonucleases / antagonists & inhibitors
  • Edetic Acid / pharmacology
  • Endopeptidase K / metabolism
  • Female
  • Fruit / enzymology
  • Genome, Human / genetics
  • Humans
  • Hydrogen-Ion Concentration / drug effects
  • Keratins / metabolism
  • Molecular Biology / methods*
  • Nails / chemistry*
  • Peptide Hydrolases / metabolism*
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Solutions
  • Temperature
  • Young Adult

Substances

  • Solutions
  • Keratins
  • DNA
  • Edetic Acid
  • Deoxyribonucleases
  • Peptide Hydrolases
  • Endopeptidase K