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      • Record: found
      • Abstract: found
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      Is Open Access

      Improved method for genomic DNA extraction for Opuntia Mill. (Cactaceae)

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          Abstract

          Background

          Genomic DNA extracted from species of Cactaceae is often contaminated with significant amounts of mucilage and pectin. Pectin is one of the main components of cellular walls, whereas mucilage is a complex polysaccharide with a ramified structure. Thus, pectin- and mucilage-free extraction of DNA is a key step for further downstream PCR-based analyses.

          Results

          We tested our DNA extraction method on cladode tissue (juvenile, adult, and herbaria exemplars) of 17 species of Opuntia Mill., which are characterized by a large quantity of pectin and mucilage.

          Conclusion

          We developed a method for the extraction of gDNA free of inhibitory compounds common in species of Opuntia Mill., such as pectin and mucilage. Compared to previously extraction protocols, our method produced higher yields of high-quality genomic DNA.

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          Most cited references40

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          Rapid isolation of high molecular weight plant DNA.

          M Murray, W. Thompson (1980)
          A method is presented for the rapid isolation of high molecular weight plant DNA (50,000 base pairs or more in length) which is free of contaminants which interfere with complete digestion by restriction endonucleases. The procedure yields total cellular DNA (i.e. nuclear, chloroplast, and mitochondrial DNA). The technique is ideal for the rapid isolation of small amounts of DNA from many different species and is also useful for large scale isolations.
          Article 0 comments Cited 950 times – based on 0 reviews     Review now
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            A simple and efficient method for DNA extraction from grapevine cultivars andVitis species

            Muhammad Lodhi, Guang-Ning Ye, Norman F. Weeden … (1994)
            Article 0 comments Cited 150 times – based on 0 reviews     Review now
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              Ribosomal RNA genes in plants: variability in copy number and in the intergenic spacer.

              Graeme Rogers, A. Bendich (1987)
              Ribosomal RNA genes in plants are highly variable both in copy number and in intergenic spacer (IGS) length. This variability exists not only between distantly related species, but among members of the same genus and also among members of the same population of a single species. Analysis of inheritance indicates that copy number change is rapid, occurring even among somatic cells of individual plants, and that up to 90% or more of the gene copies are superfluous. Subrepetitive sequences within the IGS appear to be changing rapidly as well. They are not only variable in sequence from one species to the next, but can vary in number between neighboring gene repeats on the chromosome. In all species examined in detail they are located in the same region of the IGS and contain sequences that can be folded into stem-loop structures flanked by a pyrimidine-rich region. It has been suggested that these subrepeats function in transcriptional enhancement, termination or processing, or in recombination events generating the high multiplicity of ribosomal genes.
              Article 0 comments Cited 72 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                César Ramiro Martínez-González: ramiro_mg.unam@ciencias.unam.mx
                Rosario Ramírez-Mendoza: rosaricuqui@yahoo.com.mx
                Jaime Jiménez-Ramírez: jjimenez_ramirez@yahoo.com
                Clemente Gallegos-Vázquez: cgallegosvazquez@gmail.com
                Isolda Luna-Vega: luna.isolda@gmail.com
                Journal
                Journal ID (nlm-ta): Plant Methods
                Journal ID (iso-abbrev): Plant Methods
                Title: Plant Methods
                Publisher: BioMed Central (London )
                ISSN (Electronic): 1746-4811
                Publication date (Electronic): 11 October 2017
                Publication date PMC-release: 11 October 2017
                Publication date Collection: 2017
                Volume: 13
                Electronic Location Identifier: 82
                Affiliations
                [1 ]ISNI 0000 0001 2159 0001, GRID grid.9486.3, Laboratorio de Biogeografía y Sistemática, Departamento de Biología, Facultad de Ciencias, , Universidad Nacional Autónoma de México, ; Ciudad Universitaria, Coyoacán, 04510 Mexico City, México
                [2 ]ISNI 0000 0004 1795 9752, GRID grid.418752.d, Laboratorio de Biotecnología de Semillas, , Colegio de Postgraduados, ; Carretera México-Texcoco, 56230 Estado de México, México
                [3 ]ISNI 0000 0001 2159 0001, GRID grid.9486.3, Herbario de la Facultad de Ciencias, Departamento de Biología Comparada, Facultad de Ciencias, , Universidad Nacional Autónoma de México, ; Ciudad Universitaria, Coyoacán, 04510 Mexico City, México
                [4 ]ISNI 0000 0004 0483 8492, GRID grid.34684.3d, Centro Regional Universitario Centro Norte, , Universidad Autónoma Chapingo, ; Cruz del Sur núm. 100, Colonia Constelación, El Orito, 98085 Zacatecas, Zacatecas México
                Article
                Publisher ID: 234
                DOI: 10.1186/s13007-017-0234-y
                PMC ID: 5637266
                PubMed ID: 28053646
                SO-VID: 1fddf622-313f-4790-9f52-a928c5d784cd
                Copyright © © The Author(s) 2017
                License:

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                Date received : 7 October 2016
                Date accepted : 3 October 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100005739, Universidad Nacional Autónoma de México;
                Award ID: without number
                Award Recipient :
                Funded by: Red del Nopal belonging to the Sistema Nacional de Recursos Fitogenéticos para la alimentación y la agricultura del Servicio Nacional de Inspección y certificación de semillas
                Award ID: without number
                Award Recipient :
                Categories
                Subject: Methodology
                Custom metadata
                issue-copyright-statement © The Author(s) 2017

                ScienceOpen disciplines: Plant science & Botany
                Keywords: dna quality,dna quantity,genomic dna,mucilage,opuntia,pectin
                Data availability:
                ScienceOpen disciplines: Plant science & Botany
                Keywords: dna quality, dna quantity, genomic dna, mucilage, opuntia, pectin

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