Abstract
Single-cell RNA sequencing (sc-RNAseq) has become a critical approach for the analysis of immune cell function and heterogeneity. So far, the immune cell isolation, based on surface marker expression predicted by the RNA expression profiles, is often limited by the poor correlation between transcript and protein expression patterns. To overcome these difficulties, novel single-cell multi-omic approaches based on the combined analysis of transcript and surface protein expression have been developed. One of the major benefits of these technologies is the possibility to use a high number of antibodies conjugated with oligonucleotide (AbOs) for the surface marker detection, thus overcoming the limit of using few surface markers as occurs in flow cytometry. Here we describe the BD Rhapsody single-cell analysis system protocol for 3′ mRNA whole transcriptome analysis (WTA), combined with AbO- and Sample Tag library preparation.
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BD Rhapsody™ Single-Cell Labeling with BD® Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos (41 plex to 100 plex) (Doc ID:23-22354-00)
Single Cell Capture and cDNA Synthesis with the BD Rhapsody™ Single-Cell Analysis System (Doc ID:210966 Rev. 1.0)
BD Rhapsody™ mRNA Whole Transcriptome Analysis (WTA), AbSeq, and Sample Tag Library Preparation Protocol (Doc ID: 23-21752-00)
Acknowledgments
We thank all the Armenise-Harvard Immune Regulation and IIGM members. We thank S. Scherrer for critical reading and helpful discussion. We thank E. Kowalczyk and the BD multi-omics alliance for helpful discussion, G. Granato for technical help. This work was supported by G. Armenise Harvard foundation, IIGM-CSP. L.P. has been a partner of a multi-omics alliance research program with BD (Europe). Valentina Russo and Nadia Brasu contributed equally to this work.
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Russo, V., Brasu, N., Pace, L. (2022). Combined Measurement of RNA and Protein Expression on a Single-Cell Level. In: Ooi, A.T. (eds) Single-Cell Protein Analysis. Methods in Molecular Biology, vol 2386. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1771-7_16
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DOI: https://doi.org/10.1007/978-1-0716-1771-7_16
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