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A protocol for time-resolved transcriptomics through metabolic labeling and combinatorial indexing.

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posted on 2024-10-07, 13:21 authored by Rory J Maizels, Daniel M Snell, James Briscoe
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.

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Crick (Grant ID: CC1064, Grant title: STP Advanced Sequencing) Crick (Grant ID: CC2032, Grant title: Briscoe CC2032)

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