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Mushroom DNA barcoding project: Sequencing a segment of the 28S rRNA gene.
Horn, Ivo R; Verleg, Peter A; Ibrahim, Nafiesa Z; Soeleman, Khadiedjah; van Kampen, Floris; Ruesen, Mia O; Reulen, Naïsha M; Breij, Henk; Bakker, Roderick J; Gravendeel, Barbara.
Afiliación
  • Horn IR; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • Verleg PA; Naturalis Biodiversity Center, Endless Forms Group, Leiden, The Netherlands.
  • Ibrahim NZ; Leiden Centre for Applied Bioscience, Leiden, The Netherlands.
  • Soeleman K; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • van Kampen F; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • Ruesen MO; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • Reulen NM; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • Breij H; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • Bakker RJ; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
  • Gravendeel B; University of Applied Sciences Leiden, Faculty of Science and Technology, Leiden, The Netherlands.
Biochem Mol Biol Educ ; 48(4): 404-410, 2020 07.
Article en En | MEDLINE | ID: mdl-32585770
DNA barcoding is an important molecular methodology for species identification that was developed over the last two decades and it should be covered in the biology bachelor curriculum. Here, we present an example of DNA barcoding by sequencing a segment of the 28S nuclear ribosomal large subunit rRNA gene of wild mushrooms and framing the education in a project form for undergraduate students in biology. Students perform this project in 6-8 weeks, which also includes preparing a poster, writing a report and presenting a paper related to the work in a journal club format. First, fieldwork in the Netherlands was carried out, during which students collected mushrooms under supervision of a professional mycologist with the goal to (a) verify morphologically based identifications with a molecular method and (b) assess phylogenetic relationships of the different species collected. Next, DNA extractions and quantitation were performed, PCR amplification was done, and samples were sent out for Sanger sequencing. Students aligned and analyzed the sequences using BLAST and Geneious and subsequently created a phylogenetic tree. In case of collecting DNA barcodes of an earlier sequenced species, students could upload the data to a repository established for facilitation of future research projects. The method described is very robust, reagents and equipment are readily available, and costs are relatively low. In addition, the results can be compared to published fungal phylogenetic trees.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Ribosómico 28S / Reacción en Cadena de la Polimerasa / Análisis de Secuencia de ADN / Genes de ARNr / Agaricales / Código de Barras del ADN Taxonómico Límite: Humans Idioma: En Revista: Biochem Mol Biol Educ Año: 2020 Tipo del documento: Article País de afiliación: Países Bajos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ARN Ribosómico 28S / Reacción en Cadena de la Polimerasa / Análisis de Secuencia de ADN / Genes de ARNr / Agaricales / Código de Barras del ADN Taxonómico Límite: Humans Idioma: En Revista: Biochem Mol Biol Educ Año: 2020 Tipo del documento: Article País de afiliación: Países Bajos Pais de publicación: Estados Unidos