Language
English
Publication Date
6-17-2022
Journal
STAR Protocols
DOI
10.1016/j.xpro.2022.101417
PMID
35620068
PMCID
PMC9127693
PubMedCentral® Posted Date
5-20-2022
PubMedCentral® Full Text Version
Post-print
Abstract
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 et al. (2021) and Li et al. (2022).
Keywords
Animals, Base Sequence, Drosophila, Gene Expression Profiling, Sequence Analysis, RNA, Exome Sequencing, Genomics, Model Organisms, RNAseq, Sequencing
Published Open-Access
yes
Recommended Citation
McLaughlin, Colleen N; Qi, Yanyan; Quake, Stephen R; et al., "Isolation and RNA Sequencing of Single Nuclei From Drosophila Tissues" (2022). Huffington Center on Aging Staff Publications. 54.
https://digitalcommons.library.tmc.edu/aging_research/54
Graphical Abstract