Article

Plant Molecular Biology Reporter

, Volume 16, Issue 1, pp 69-86

First online:

Comparative Analysis of Different DNA Extraction Protocols: A Fast, Universal Maxi-Preparation of High Quality Plant DNA for Genetic Evaluation and Phylogenetic Studies

  • U.M. CsaiklAffiliated withAustrian Research Centre Seibersdorf
  • , H. BastianAffiliated withQIAGEN GmbH
  • , R. BrettschneiderAffiliated withCentre for Applied Plant Molecular Biology, AMPII, Institute of General Botany
  • , S. GauchAffiliated withQIAGEN GmbH
  • , A. MeirAffiliated withQIAGEN GmbH
  • , M. SchauerteAffiliated withFederal Research Centre for Forestry and Forest Products, Institute for Forest Genetics
  • , F. ScholzAffiliated withFederal Research Centre for Forestry and Forest Products, Institute for Forest Genetics
  • , C. SperisenAffiliated withSnow and Landscape Research, Swiss Federal Institute for Forest
  • , B. VornamAffiliated withInstitute for Forest Genetics and Plant Breeding, University of Göttingen
    • , B. ZiegenhagenAffiliated withFederal Research Centre for Forestry and Forest Products, Institute for Forest Genetics

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Abstract

Four DNA extraction protocols were compared for ability to produce DNA from the leaves or needles of several species: oak, elm, pine, fir, poplar and maize (fresh materials) and rhododendron (silica dried or frozen material). With the exception of maize and poplar, the species are known to be difficult for DNA extraction. Two protocols represented classical procedures for lysis and purification, and the other two were a combination of classical lysis followed by anion exchange chromatography. The DNA obtained from all procedures was quantified and tested by PCR and Southern hybridisation.Test results indicated superiority of one of the four protocols; a combination of CTAB lysis followed by anion exchange chromatography which enabled DNA extraction from all seven species. A second protocol also produced DNA from leaves or needles of all species investigated and was well suited for PCR applications but not Southern hybridisations. The remaining protocols produced DNA from some but not all species tested.

Abbreviations: CTAB, hexadecyltrimethylammonium bromide; EtOH, Ethanol; TBE, tris-borate-EDTA.

cpDNA DNA extraction fingerprinting forest trees M13 fingerprinting method PCR rDNA RFLP rhododendron plant