Preparation of Drosophila Ovarioles for Single-Cell RNA Sequencing.
Preparation of Drosophila Ovarioles for Single-Cell RNA Sequencing.
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用于单细胞 RNA 测序的果蝇卵巢的制备。
DOI：
10.1007/978-1-0716-2970-3_17
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Nystul,Todd
中科院分区：
文献类型：
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作者：
Meyer,Nathaniel;Peralta,Jobelle;Nystul,Todd
在果蝇卵巢中产生卵子需要共存于同一固体组织中的多种细胞类型之间复杂的相互作用。这种细胞的异质性使卵巢成为一个丰富的研究主题，但也使使用批量RNA测序等方法识别单个细胞类型之间的转录差异具有挑战性。单细胞RNA测序(scRNA-seq)技术的发展提供了一种在细胞分辨率下分析遗传和功能异质性的途径，从而解决了这一限制。在这里，我们描述了果蝇卵巢的分离和制备以用于scRNA-seq。该方案强调准备时间短，细胞活力高，防止RNA降解，减少技术变异，以实现高度可重复性的单细胞图谱。
The production of eggs in theDrosophilaovary requires complex interactions between multiple cell types that coexist within the same solid tissue. This cellular heterogeneity makes the ovary a rich subject of study, but also makes it challenging to identify transcriptional differences between individual cell types using methods such as bulk RNA sequencing. The development of single-cell RNA sequencing (scRNA-seq) techniques addresses this limitation by providing an avenue to profile genetic and functional heterogeneity at a cellular resolution. Here, we describe the isolation and preparation of theDrosophilaovary for scRNA-seq. This protocol emphasizes a short preparation time, high cell viability, prevention of RNA-degradation, and reduction of technical variation to achieve highly reproducible single-cell profiles.