High-throughput scNMT protocol for multiomics profiling of single cells from mouse brain and pancreatic organoids

Summary: Single-cell nucleosome, methylome, and transcriptome (scNMT) sequencing is a recently developed method that allows multiomics profiling of single cells. In this scNMT protocol, we describe profiling of cells from mouse brain and pancreatic organoids, using liquid handling platforms to incre...

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Bibliographic Details
Main Authors: Santiago Cerrizuela, Oguzhan Kaya, Lukas P.M. Kremer, Andrea Sarvari, Tobias Ellinger, Jannes Straub, Jan Brunken, Andrés Sanz-Morejón, Aylin Korkmaz, Ana Martín-Villalba
Format: Article
Language:English
Published: Elsevier 2022-09-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S266616672200435X
Description
Summary:Summary: Single-cell nucleosome, methylome, and transcriptome (scNMT) sequencing is a recently developed method that allows multiomics profiling of single cells. In this scNMT protocol, we describe profiling of cells from mouse brain and pancreatic organoids, using liquid handling platforms to increase throughput from 96-well to 384-well plate format. Our approach miniaturizes reaction volumes and incorporates the latest Smart-seq3 protocol to obtain higher numbers of detected genes and genomic DNA (gDNA) CpGs per cell. We outline normalization steps to optimally distribute per-cell sequencing depth.For complete details on the use and execution of this protocol, please refer to Clark (2019), Clark et al. (2018), and Hagemann-Jensen et al. (2020a, 2020b). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667