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RNA-free DNA Extraction Protocol from PinusTissues for Molecular Biology or HPCE/HPLC Analyses

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Abstract

A simple method for extracting DNA from various in vitro or ex vitro Pinus tissues is described. Good yield and high quality RNA-free DNA ready to use in molecular biology assays or analytical analysis was obtained. DNA quality was measured analyzing absorption spectra between 200 and 300 nm, A260/280 ratio and HPCE. This protocol was tested with no modification in a wide range of Pinus tissues from recalcitrant in vitro callus to mature field scions improving the results obtained with previous protocols. As the protocol is simple, almost universal and inexpensive it may be used for routine isolation from Pinus tissues. An isolation troubleshooting table was included to facilitate the setting up of reported protocol to other plant species.

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Abbreviations

CTAB:

Cetrimonium bromide

DIECA:

diethyldithiocarbamic acid

EDTA:

Ethylenediaminetetraacetic acid

HPCE:

High performance capillary electrophoresis

HPLC:

High performance liquid chromatography

PVP:

Polyvinylpyrrolidone

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Authors and Affiliations

  1. EPIPHYSAGE Research Group, Plant Physiology, BOS Department and IUBA, University of Oviedo, Oviedo, Spain

    L. Valledor, R. Rodríguez & M. J. Cañal

  2. Genómica Forestal S.A. Biotechnology Centre, University of Concepción, Concepción, Chile

    R. Hasbún

Authors
  1. L. Valledor
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  2. R. Hasbún
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  3. R. Rodríguez
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  4. M. J. Cañal
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Corresponding author

Correspondence to M. J. Cañal.

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Valledor, L., Hasbún, R., Rodríguez, R. et al. RNA-free DNA Extraction Protocol from PinusTissues for Molecular Biology or HPCE/HPLC Analyses. J. Plant Biochem. Biotechnol. 18, 229–232 (2009). https://doi.org/10.1007/BF03263325

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  • DOI: https://doi.org/10.1007/BF03263325

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