Robust CTAB-activated charcoal protocol for plant DNA extraction

Authors

  • Mitja KRIŽMAN SI-1000 Ljubljana, Hajdrihova 19, Slovenia
  • Jernej JAKŠE Biotechnical Faculty, University of Ljubljana, SI-1000 Ljubljana Jamnikarjeva 101, Slovenia
  • Dea BARIČEVIČ SI-1000 Ljubljana Jamnikarjeva 101, Slovenia
  • Branka JAVORNIK SI-1000 Ljubljana Jamnikarjeva 101, Slovenia
  • Mirko PROŠEK SI-1000 Ljubljana, Hajdrihova 19, Slovenia

DOI:

https://doi.org/10.14720/aas.2006.87.2.15122

Keywords:

activated charcoal, DNA extraction, PCR amplification, polyvinylpyrrolidone

Abstract

DNA extracted from plants rich in polyphenols and/or polysaccharides is often problematic when subjected to polymerase chain reaction, especially when mature tissues are used for DNA extraction. In order to overcome the problems associated with poor-quality DNA extracted from such plant samples, a protocol has been developed, availing on a high salt concentration and on the combination of polyvinylpyrrolidone and activated charcoal in the extraction buffer, in order to prevent the solubilization of polysaccharides and polyphenols in the DNA extract. Mild temperature conditions during extraction and precipitation were also recognized as important parameters. Besides DNA purity, mild precipitation conditions were found to be beneficial in obtaining less low-molecular mass nucleic acids in the final DNA extract. The homogenization step and the amount of sample extracted were also found to be crucial in keeping the extraction procedure robust.

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Published

15. 09. 2006

Issue

Section

Original Scientific Article

How to Cite

KRIŽMAN, M., JAKŠE, J., BARIČEVIČ, D., JAVORNIK, B., & PROŠEK, M. (2006). Robust CTAB-activated charcoal protocol for plant DNA extraction. Acta Agriculturae Slovenica, 87(2), 427–433. https://doi.org/10.14720/aas.2006.87.2.15122

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