Synthesis and application of POLYseq for profiling human liver organoids

STAR Protoc. 2021 Dec 4;2(4):100976. doi: 10.1016/j.xpro.2021.100976. eCollection 2021 Dec 17.

Abstract

Detailed herein is the protocol for synthesis, characterization, and application of POLYseq for cell pooling in single-cell sequencing runs. POLYseq is synthesized through commercially available reagents and is highly tailorable. Synthesis is easily performed in a two-step protocol, utilizing Michael addition only requiring a temperature-stable hot bath capable of holding 90°C. However, care must be taken when mixing reagents for synthesis, as the final product is sensitive to initial mixing ratios of POLYseq reagents. For complete details on the use and execution of this protocol, please refer to Dunn et al. (2021).

Keywords: Biotechnology and bioengineering; Cell-based Assays; Chemistry; Organoids; RNAseq; Sequencing; Single Cell; Stem Cells.

Publication types

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

MeSH terms

  • Humans
  • Liver / cytology*
  • Optical Imaging / methods*
  • Organoids / cytology*
  • Sequence Analysis, DNA / methods*
  • Single-Cell Analysis / methods*