方案更新为：用于小鼠脑和胰腺类器官单细胞多组学分析的高通量scNMT方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-07-24 DOI:10.1016/j.xpro.2025.103980

Santiago Cerrizuela, Oguzhan Kaya, Lukas P M Kremer, Andrea Sarvari, Tobias Ellinger, Jannes Straub, Jan Brunken, Andrés Sanz-Morejón, Aylin Korkmaz, Ana Martín-Villalba

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引用次数: 0

摘要

单细胞核小体、甲基组和转录组（scNMT）测序是最近发展起来的一种方法，可以对单细胞进行多组学分析。在本scNMT协议中，我们描述了来自小鼠大脑和胰腺类器官的细胞谱，使用液体处理平台将通量从96孔增加到384孔板格式。我们的方法使反应体积小型化，并结合了最新的Smart-seq3协议，以获得每个细胞更高数量的检测基因和基因组DNA (gDNA) CpGs。我们概述了标准化的步骤，以最佳地分配每细胞测序深度。关于本协议使用和执行的完整细节，请参考Kremer等人的著作。1、2、3、4、5、6、7该协议是对Cerrizuela等人的更新。

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Protocol update to: 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 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 Kremer et al. and other works.¹^,²^,³^,⁴^,⁵^,⁶^,⁷ This protocol is an update to Cerrizuela et al.⁷.