Whole genome amplification and sequencing of a Daphnia resting egg

Justin B. Lack, Lawrence J. Weider, Punidan D. Jeyasingh

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Resting eggs banks are unique windows that allow us to directly observe shifts in population genetics, and phenotypes over time as natural populations evolve. Though a variety of planktonic organisms also produce resting stages, the keystone freshwater consumer, Daphnia, is a well-known model for paleogenetics and resurrection ecology. Nevertheless, paleogenomic investigations are limited largely because resting eggs do not contain enough DNA for genomic sequencing. In fact, genomic studies even on extant populations include a laborious preparatory phase of batch culturing dozens of individuals to generate sufficient genomic DNA. Here, we furnish a protocol to generate whole genomes of single ephippial (resting) eggs and single daphniids. Whole genomes of single ephippial eggs and single adults were amplified using Qiagen REPLI-g Single Cell kit reaction, followed by NEBNext Ultra DNA Library Prep Kit for library construction and Illumina sequencing. We compared the quality of the single-egg and single-individual amplified genomes to the standard batch genomic DNA extraction in the absence of genome amplification. At mean 20× depth, coverage was essentially identical for the amplified single individual relative to the unamplified batch extracted genome (>90% of the genome was covered and callable). Finally, while amplification resulted in the slight loss of heterozygosity for the amplified genomes, estimates were largely comparable and illustrate the utility and limitations of this approach in estimating population genetic parameters over long periods of time in natural populations of Daphnia and also other small species known to produce resting stages.

Original languageEnglish (US)
Pages (from-to)118-127
Number of pages10
JournalMolecular Ecology Resources
Volume18
Issue number1
DOIs
StatePublished - Jan 2018

Bibliographical note

Funding Information:
National Science Foundation, Grant/Award Number: 1256867 and 1256881

Funding Information:
We are grateful to the UW-Madison Center for high-throughput computing for providing computational assistance and sequencing resources. We thank Aime’e Yousey for laboratory assistance. NSF Grant # 1256867 (P.D.J.) and #1256881 (L.J.W.) supported this work. We thank the editor and three anonymous reviewers for constructive comments.

Publisher Copyright:
© 2017 John Wiley & Sons Ltd

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

Keywords

  • ancient DNA
  • egg bank
  • ephippia
  • paleogenomics
  • resurrection ecology
  • zooplankton

Continental Scientific Drilling Facility tags

  • DAPH

Fingerprint

Dive into the research topics of 'Whole genome amplification and sequencing of a Daphnia resting egg'. Together they form a unique fingerprint.

Cite this