scMTR-seq
Abstract
Here, we describe a protocol for scMTR-seq (single cell multi-targets and mRNA sequencing), which is a multi-omics sequencing technology that can simultaneously profile multiple histone modifications or other chromatin-bound proteins and transcriptome in the same single cells. scMTR-seq has high sensitivity, high cell recovery and is highly scalable in terms of starting material requirement. The protocol has the following steps: i) pre-assemble antibodies specific for each target histone modification or protein with indexed proteinA-Tn5-adapters; ii) perform in situ Tn5-mediated tagmentation with indexed complexes; iii) capture nuclear mRNA with a barcoded poly-T primer followed by in situ reverse transcription; iv) label single cells using split-pool combinatorial barcoding. We have applied scMTR-seq to profile histone modifications and transcriptomes in single cells from a broad range of heterogeneous samples including pluripotent stem cell differentiation and mouse pre-implantation embryos.
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DOI:10.17504/protocols.io.yxmvmb7dog3p/v1