Optimized Replication of Arrayed Bacterial Mutant Libraries Increases Access to Biological Resources

Julia L.E. Willett, Aaron M.T. Barnes, Debra N. Brunson, Alexandre Lecomte, Ethan B. Robertson, Gary M. Dunny

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Biological collections, including arrayed libraries of single transposon (Tn) or deletion mutants, greatly accelerate the pace of bacterial genetic research. Despite the importance of these resources, few protocols exist for the replication and distribution of these materials. Here, we describe a protocol for creating multiple replicates of an arrayed bacterial Tn library consisting of approximately 6,800 mutants in 96-well plates (73 plates). Our protocol provides multiple checkpoints to guard against contamination and minimize genetic drift caused by freeze/thaw cycles. This approach can also be scaled for arrayed culture collections of various sizes. Overall, this protocol is a valuable resource for other researchers considering the construction and distribution of arrayed culture collection resources for the benefit of the greater scientific community.

Original languageEnglish (US)
JournalMicrobiology Spectrum
Volume11
Issue number4
DOIs
StatePublished - Aug 2023

Bibliographical note

Publisher Copyright:
© 2023 Willett et al.

Keywords

  • Enterococcus faecalis
  • biological collections
  • functional genomics
  • genetic tools
  • scientific rigor
  • transposon mutagenesis

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

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