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
| Autor: | Marissa E Klingseis, Danielle R Edelman, Reid N Brown, Bethany Bolander, Steven R. Bilby, Christina E Gray, Amie G Haddock, Rachael A Prawitz, Alexander M. Duryee, Mackenzie M Helsel, Jeremy H Brown, Edward W Miles, Kalif Leslie, Chandler Gossett, Alyssa D Jones, Csengele Barta |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Plant Science Isoprene synthase Secondary metabolite 01 natural sciences isoprene synthase (IspS) Article law.invention 03 medical and health sciences chemistry.chemical_compound law lcsh:Botany Botany medicine Gene DNA extraction pin oak (Quercus palustris) Ecology Evolution Behavior and Systematics Quercus palustris Polymerase chain reaction gene identification Ecology biology secondary metabolites leaves dark adaptation PCR 18S rRNA biology.organism_classification lcsh:QK1-989 genomic DNA 030104 developmental biology chemistry biology.protein DNA 010606 plant biology & botany medicine.drug |
| Zdroj: | Plants; Volume 6; Issue 4; Pages: 52 Plants, Vol 6, Iss 4, p 52 (2017) Plants |
| ISSN: | 2223-7747 |
| DOI: | 10.3390/plants6040052 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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