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

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 throu...

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Detalles Bibliográficos
Autores principales: Cerrizuela, Santiago, Kaya, Oguzhan, Kremer, Lukas P.M., Sarvari, Andrea, Ellinger, Tobias, Straub, Jannes, Brunken, Jan, Sanz-Morejón, Andrés, Korkmaz, Aylin, Martín-Villalba, Ana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9441340/
https://www.ncbi.nlm.nih.gov/pubmed/36072757
http://dx.doi.org/10.1016/j.xpro.2022.101555
Descripción
Sumario: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 Clark et al., 2018, Hagemann-Jensen et al., 2020a, Hagemann-Jensen et al., 2020b.

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