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Semi-automated, Membrane-Based Protocol for DNA Isolation from Plants

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Abstract

Many plant species are considered difficult for DNA isolation due to their high concentrations of secondary metabolites such as polysaccharides and polyphenols. Several protocols have been developed to overcome this problem, but they are typically time-consuming, not scalable for high throughput and not compatible with automation. Although a variety of commercial kits are available for plant DNA isolation, their cost is high and these kits usually have limited taxonomic applicability. In a previous study we developed an inexpensive automation-friendly protocol for DNA extraction from animal tissues. Here we demonstrate that a similar protocol allows DNA isolation from plants.

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Notes

  1. To reduce the probability of cross-contamination due to airborne plant material, place one strip of tubes in a separate rack during sampling and after homogenization. After homogenization open tubes carefully using the individual side tabs of each tube. Discard the lids, replace with new ones after addition of lysis buffer and return the strip to original rack.

Abbreviations

CTAB:

cetyltrimethylammonium bromide

PVP:

polyvinylpyrrolidone

ILB:

insect lysis buffer

GuSCN:

guanidine thiocyanate

SDS:

sodium dodecyl sulfate

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Acknowledgements

This work was supported by grants to PDNH from Genome Canada through the Ontario Genomics Institute, the Canada Foundation for Innovation, the Ontario Innovation Trust, the Canada Research Chairs Program and NSERC. We thank M. Hajibabaei for comments on the manuscript, J. Gerrath for assistance in the field and I. Meusier for evaluation of the automated protocol.

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Authors and Affiliations

  1. Canadian Centre for DNA Barcoding, Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, N1G 2W1, Canada

    Natalia V. Ivanova & Paul D. N. Hebert

  2. Department of Integrative Biology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada

    Aron J. Fazekas

Authors
  1. Natalia V. Ivanova
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  2. Aron J. Fazekas
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  3. Paul D. N. Hebert
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Correspondence to Natalia V. Ivanova.

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Ivanova, N.V., Fazekas, A.J. & Hebert, P.D.N. Semi-automated, Membrane-Based Protocol for DNA Isolation from Plants. Plant Mol Biol Rep 26, 186–198 (2008). https://doi.org/10.1007/s11105-008-0029-4

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