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Plants 2017, 6(4), 52; https://doi.org/10.3390/plants6040052

In Situ Dark Adaptation Enhances the Efficiency of DNA Extraction from Mature Pin Oak (Quercus palustris) Leaves, Facilitating the Identification of Partial Sequences of the 18S rRNA and Isoprene Synthase (IspS) Genes

Department of Biology, Missouri Western State University, 4525 Downs Drive, Agenstein-Remington Halls, St. Joseph, MO 64507, USA
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Received: 28 August 2017 / Revised: 29 September 2017 / Accepted: 19 October 2017 / Published: 24 October 2017
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Abstract

Mature oak (Quercus spp.) leaves, although abundantly available during the plants’ developmental cycle, are rarely exploited as viable sources of genomic DNA. These leaves are rich in metabolites difficult to remove during standard DNA purification, interfering with downstream molecular genetics applications. The current work assessed whether in situ dark adaptation, to deplete sugar reserves and inhibit secondary metabolite synthesis could compensate for the difficulties encountered when isolating DNA from mature leaves rich in secondary metabolites. We optimized a rapid, commercial kit based method to extract genomic DNA from dark- and light-adapted leaves. We demonstrated that in situ dark adaptation increases the yield and quality of genomic DNA obtained from mature oak leaves, yielding templates of sufficiently high quality for direct downstream applications, such as PCR amplification and gene identification. The quality of templates isolated from dark-adapted pin oak leaves particularly improved the amplification of larger fragments in our experiments. From DNA extracts prepared with our optimized method, we identified for the first time partial segments of the genes encoding 18S rRNA and isoprene synthase (IspS) from pin oak (Quercus palustris), whose full genome has not yet been sequenced. View Full-Text
Keywords: DNA extraction; pin oak (Quercus palustris); leaves; dark adaptation; secondary metabolites; PCR; gene identification; 18S rRNA; isoprene synthase (IspS) DNA extraction; pin oak (Quercus palustris); leaves; dark adaptation; secondary metabolites; PCR; gene identification; 18S rRNA; isoprene synthase (IspS)
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Barta, C.E.; Bolander, B.; Bilby, S.R.; Brown, J.H.; Brown, R.N.; Duryee, A.M.; Edelman, D.R.; Gray, C.E.; Gossett, C.; Haddock, A.G.; Helsel, M.M.; Jones, A.D.; Klingseis, M.E.; Leslie, K.; Miles, E.W.; Prawitz, R.A. In Situ Dark Adaptation Enhances the Efficiency of DNA Extraction from Mature Pin Oak (Quercus palustris) Leaves, Facilitating the Identification of Partial Sequences of the 18S rRNA and Isoprene Synthase (IspS) Genes. Plants 2017, 6, 52.

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