Comparison of methods of DNA extraction from herbarium specimens of little-pod false flax (Camelina microcarpa Andrz. Ex Dc.)

  • V. H. Sakharova Institute of Food Biotechnology and Genomics, Nat. Acad. Sci. of Ukraine, Ukraine, 04123, Kyiv, Osypovskogo str., 2a
  • R. Ya. Blume Institute of Food Biotechnology and Genomics, Nat. Acad. Sci. of Ukraine, Ukraine, 04123, Kyiv, Osypovskogo str., 2a; Educational and Scientific Center "Institute of Biology and Medicine", Taras Shevchenko National University of Kyiv, Ukraine, 03022, Kyiv, Academika Glushkova ave., 2
  • A. M. Rabokon nstitute of Food Biotechnology and Genomics, Nat. Acad. of Sci. of Ukraine, Ukraine, 04123, Kyiv, Оsypovskoho str., 2A https://orcid.org/0000-0002-6249-1824
  • Ya. V. Pirko Institute of Food Biotechnology and Genomics, Nat. Acad. of Sci. of Ukraine, Ukraine, 04123, Kyiv, Оsypovskoho str., 2A https://orcid.org/0000-0003-1887-5406
  • Ya. B. Blume Institute of Food Biotechnology and Genomics, Nat. Acad. of Sci. of Ukraine, Ukraine, 04123, Kyiv, Оsypovskoho str., 2A https://orcid.org/0000-0001-7078-7548
DOI: https://doi.org/10.7124/FEEO.v30.1457
Keywords: Camelina microcarpa Andrz. ex DC., little-pod false flax, DNA isolation, herbarium specimens

Abstract

Aim. The aim of this research was to compare the efficiency of DNA isolation methods from herbarium specimens of Camelina microcarpa Andrz. Ex DC., further modification of these methods to increase DNA yield, and determine the method that would provide the best yield of isolated DNA. Methods. Modifications of the DNA isolation methods using the DNeasy Plant Mini Kit (QIAgen) and the CTAB method were used. PCR was performed using degenerate primers for method of β-tubulin intron length polymorphism (TBP). Amplicons were fractionated in polyacrylamide gel followed by visualization by silver nitrate staining. Results. DNA was successfully extracted from C. microcarpa herbarium specimens sampled with leaf parts and seeds, using the modified by CTAB method, and four modified methods using DNeasy Plant Mini Kit (QIAgen). Conclusions. The study revealed that the most effective method tested was the DNeasy Plant Mini Kit (QIAgen) No. 2. Prolongation of the cell lysis stage had the best effect on the increase of DNA yield. We found that the success of DNA isolation was influenced not so much by the age of the herbarium specimen as by the methods of drying and storing the plants in the collection.

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