STAR Protocols (Mar 2023)

Multiplexed evaluation of mouse wound tissue using oligonucleotide barcoding with single-cell RNA sequencing

  • Michael Januszyk,
  • Michelle Griffin,
  • Shamik Mascharak,
  • Heather E. Talbott,
  • Kellen Chen,
  • Dominic Henn,
  • Amanda F. Spielman,
  • Jennifer B.L. Parker,
  • Norah E. Liang,
  • Asha Cotterell,
  • Nicholas Guardino,
  • Deshka S. Foster,
  • Dhananjay Wagh,
  • John Coller,
  • Geoffrey C. Gurtner,
  • Derrick C. Wan,
  • Michael T. Longaker

Journal volume & issue
Vol. 4, no. 1
p. 101946

Abstract

Read online

Summary: Despite its rapidly increased availability for the study of complex tissue, single-cell RNA sequencing remains prohibitively expensive for large studies. Here, we present a protocol using oligonucleotide barcoding for the tagging and pooling of multiple samples from healing wounds, which are among the most challenging tissue types for this application. We describe steps to generate skin wounds in mice, followed by tissue harvest and oligonucleotide barcoding. This protocol is also applicable to other species including rats, pigs, and humans.For complete details on the use and execution of this protocol, please refer to Stoeckius et al. (2018),1 Galiano et al. (2004),2 and Mascharak et al. (2022).3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords