Abstract
Transcriptome profiling has been significantly hampered by the heterogeneity among individual cells within a tissue or an organ. Recent advances in single cell transcriptome profiling have significantly advanced our understanding of the transcriptome. However, plant single-cell RNA sequencing (scRNA-seq) relies on the isolation of protoplasts, which is not only impossible for many cell types but also induces acute wounding responses. To solve these problems, single-nucleus RNA sequencing (snRNA-seq) has been applied to plant research, in which nuclei are isolated and subject to encapsulation and profiling. Compared with scRNA-seq, snRNA-seq can be applied to a wider range of tissue types and plant species. Nevertheless, fewer transcripts can be obtained from each nucleus than each protoplast. In this chapter, we describe a detailed and general protocol to prepare nuclei from plant tissues that are ready for subsequent library construction and high-throughput sequencing.
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Acknowledgments
This work was supported by the CAS Strategic Priority Research Program (grant XDA24020203) and the National Key R&D Program of China (grant 2023YFE0101100).
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Xin, X., Du, F., Jiao, Y. (2023). Plant Nuclei Isolation for Single-Nucleus RNA Sequencing. In: Riechmann, J.L., Ferrándiz, C. (eds) Flower Development . Methods in Molecular Biology, vol 2686. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3299-4_15
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DOI: https://doi.org/10.1007/978-1-0716-3299-4_15
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