A protocol for time-resolved transcriptomics through metabolic labeling and combinatorial indexing
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The snapshot nature of single-cell transcriptomics presents a challenge for studying the dynamics of gene expression. Metabolic labeling, where nascent RNA is labeled with 4-thiouridine (4sU), captures temporal information at the single-cell level, providing greater insight into expression dynamics. Here, we present an optimized, automation-friendly protocol for the metabolic labeling of RNA alongside single-cell RNA sequencing through combinatorial indexing. We describe steps for 4sU labeling, cell fixation and chemical treatment, and automated two-level combinatorial indexing. For complete details on the use and execution of this protocol, please refer to Maizels et al.1.
Journal details
Journal
STAR Protocols
Volume
5
Issue number
4
Pages
103356
Available online
Publication date
Full text links
Publisher website (DOI)
10.1016/j.xpro.2024.103356
Europe PubMed Central
39356643
Pubmed
39356643
