SAG-RAD: A Method for Single-Cell Population Genomics of Unicellular Eukaryotes

Mol Biol Evol. 2023 May 2;40(5):msad095. doi: 10.1093/molbev/msad095.


Sequencing of reduced representation libraries enables genotyping of many individuals for population genomic studies. However, high amounts of DNA are required, and the method cannot be applied directly on single cells, preventing its use on most microbes. We developed and implemented the analysis of single amplified genomes followed by restriction-site-associated DNA sequencing to bypass labor-intensive culturing and to avoid culturing bias in population genomic studies of unicellular eukaryotes. This method thus opens the way for addressing important questions about the genetic diversity, gene flow, adaptation, dispersal, and biogeography of hitherto unexplored species.

Keywords: culturing bias; microbial genetic diversity; microeukaryotes; restriction-site associated DNA (RAD) sequencing; single amplified genome (SAG); whole-genome amplification (WGA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Eukaryota* / genetics
  • Genome
  • Genomics / methods
  • Metagenomics*
  • Sequence Analysis, DNA / methods