Protocol to achieve high-resolution single-cell transcriptomics of cardiomyocytes in multiple species

Autor: Ditte Gry Ellman, Frederik Adam Bjerre, Sara Thornby Bak, Sabrina Bech Mathiesen, Eva Bang Harvald, Charlotte Harken Jensen, Ditte Caroline Andersen
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: STAR Protocols, Vol 5, Iss 3, Pp 103194- (2024)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2024.103194
Popis: Summary: Single-cell RNA sequencing (scRNA-seq) remains state-of-the-art for transcriptomic cell-mapping. Here, we provide a protocol to generate high-resolution scRNA-seq of rare cardiomyocyte populations (e.g., regenerating/dividing, etc.) from mouse and zebrafish hearts as well as induced pluripotent stem cells, collected in time to achieve detailed transcriptomic insight. We describe the serial steps of viability staining, methanol fixation, storage, and cell sorting to preserve RNA integrity suited for scRNA-seq as well as the quality assessment of the data as shown by examples.For complete details on the use and execution of this protocol, please refer to Bak et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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