DNA Barcoding in a Field Setting v1

Résumé

This protocol was used in the 2018 Genomics in the Jungle field course held at the Inkaterra Green Lab in the Madre de Dios Department of Peru. We were able to use it to take 112 amplicons and multiplex them using a 96 barcode kit on an Oxford Nanopore Technologies MinION sequencer. This protocol can be used to multiplex much larger numbers of samples onto a single flowcell if the amplicons are from differing taxonomic groups. We multiplexed the following taxonomic groups: 1. Invertebrates 2. Mammals 3. Plants 4. Environmental DNA We isolated amplicons using markers for: 1. rDNA (see Krehenwinkel, H., Pomerantz, A., Henderson, J.B., Kennedy, S.R., Lim, J.Y., Swamy, V., Shoobridge, J.D., Patel, N.H., Gillespie, R.G. and Prost, S., 2018. Nanopore sequencing of long ribosomal DNA amplicons enables portable and simple biodiversity assessments with high phylogenetic resolution across broad taxonomic scale.bioRxiv, p.358572.) 2. COI using a mammal cocktail as recommended in Kress, W.J. and Erickson, D.L., 2012. DNA barcodes: methods and protocols. InDNA Barcodes(pp. 3-8). Humana Press, Totowa, NJ. 3. rBCL and matK for plant samples

Citer ce document

Accès au document

Ce lien n'est plus accessible actuellement. Contactez l'institution d'origine.

Auteur(s)

Statistiques

Consultations : 1

Téléchargements : 0