STAR protocols | VOL. 3

Isolation and RNA sequencing of single nuclei fromDrosophila tissues.

Publication Date Jun 17, 2022


Many insect cells are encapsulated within the exoskeleton and cannot be dissociated intact, making them inaccessible to single-cell transcriptomic profiling. We have used single-nucleus RNA sequencing to extract transcriptomic information from multiple Drosophila tissues. Here, we describe procedures for the (1) dissociation of single nuclei, (2) isolation of single nuclei using two popular cell sorters, and (3) preparation of libraries for Smart-seq2 and 10× Genomics. This protocol enables generation of high-quality transcriptomes from single nuclei and can be applied to other species. For complete details on the use and execution of this protocol, please refer to McLaughlin etal. (2021) and Li etal. (2022).


RNA Sequencing Of Tissues Single Nuclei Insect Cells Procedures For Dissociation Isolation Of Nuclei RNA Sequencing Single-cell Profiling Single Tissues Single-nucleus RNA Sequencing Single-cell Transcriptomic Profiling

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