Protocol for nuclear dissociation of the adult C. elegans for single-nucleus RNA sequencing and its application for mapping environmental responses.
Protocol for nuclear dissociation of the adult C. elegans for single-nucleus RNA sequencing and its application for mapping environmental responses.
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线虫成虫核解离的单核RNA测序方法及其在绘制环境响应图中的应用。
DOI：
10.1016/j.xpro.2023.102756
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发表时间：
2023-12-15
期刊：
影响因子：
--
通讯作者：
Allard, Patrick
中科院分区：
文献类型：
--
作者：
Levenson, Max T.;Barrere-Cain, Rio;Truong, Lisa;Chen, Yen-Wei;Shuck, Karissa;Panter, Blake;Reich, Ella;Yang, Xia;Allard, Patrick
秀丽隐杆线虫是研究器官、组织和细胞对外界刺激反应的重要模型。然而，线虫包含多个合胞体组织，其空间坐标对应于不同的核转录组。在这里，我们提出了一个单核RNA测序（snRNA-seq）协议，旨在克服在秀丽隐杆线虫单细胞RNA测序遇到的困难。我们描述了分离秀丽隐杆线虫细胞核用于下游应用的步骤，包括应用于酒精暴露背景的snRNA-seq。有关本方案使用和执行的完整详细信息，请参见Truong等人。（2023年）。秀丽隐杆线虫的全身核解离用于单核RNA测序秀丽隐杆线虫酒精液体暴露用于snRNA-seq应用的详细步骤蠕虫生长条件，以获得P0暴露后的F1和F3代出版者注：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。秀丽隐杆线虫是研究器官、组织和细胞对外界刺激反应的重要模型。然而，线虫包含多个合胞体组织，其空间坐标对应于不同的核转录组。在这里，我们提出了一个单核RNA测序（snRNA-seq）协议，旨在克服在秀丽隐杆线虫单细胞RNA测序遇到的困难。我们描述了分离秀丽隐杆线虫细胞核用于下游应用的步骤，包括应用于酒精暴露背景的snRNA-seq。
Caenorhabditis elegans is a valuable model to study organ, tissue, and cell-type responses to external cues. However, the nematode comprises multiple syncytial tissues with spatial coordinates corresponding to distinct nuclear transcriptomes. Here, we present a single-nucleus RNA sequencing (snRNA-seq) protocol that aims to overcome difficulties encountered with single-cell RNA sequencing in C. elegans. We describe steps for isolating C. elegans nuclei for downstream applications including snRNA-seq applied to the context of alcohol exposure. For complete details on the use and execution of this protocol, please refer to Truong et al. (2023). Whole-body nuclear dissociation of C. elegans for single-nucleus RNA sequencing Detailed steps for C. elegans alcohol liquid exposure for snRNA-seq applications Worm growth conditions to obtain the F1 and F3 generations after exposure in the P0 Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Caenorhabditis elegans is a valuable model to study organ, tissue, and cell-type responses to external cues. However, the nematode comprises multiple syncytial tissues with spatial coordinates corresponding to distinct nuclear transcriptomes. Here, we present a single-nucleus RNA sequencing (snRNA-seq) protocol that aims to overcome difficulties encountered with single-cell RNA sequencing in C. elegans. We describe steps for isolating C. elegans nuclei for downstream applications including snRNA-seq applied to the context of alcohol exposure.
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