A low-cost platform suitable for sequencing-based recovery of natural variation in understudied plants

Biotechniques. 2021 Feb;70(2):120-125. doi: 10.2144/btn-2020-0132. Epub 2020 Dec 14.

Abstract

Genetic characterization of wild and cultivated plants provides valuable knowledge for conservation and agriculture. DNA sequencing technologies are improving, and costs are dropping. Yet analysis of many species is hindered because they grow in regions that lack infrastructure for advanced molecular biology. The authors developed and adapted low-cost methods that address these issues. Tissue was collected and stored in silica gel, avoiding the need for liquid nitrogen and freezers. The authors optimized low-cost, homemade DNA extraction to increase yields, reduce costs and produce DNA suitable for next-generation sequencing. The authors describe how to build a gel documentation system for DNA quantification. As a proof of principle, the authors used these methods to evaluate wild Berberis darwinii, native to Southern Chile.

Keywords: Berberis darwinii; DNA extraction; do-it-yourself; gel documentation; genotyping-by-sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Berberis / genetics
  • Chile
  • DNA
  • Genetic Variation*
  • High-Throughput Nucleotide Sequencing*
  • Plants* / genetics
  • Sequence Analysis, DNA

Substances

  • DNA