Multimodal chromatin profiling using nanobody-based single-cell CUT&Tag
). The number of cells used as input was 200,000. The cells were counted, centrifuged for 10 min at 300, resuspended at 4 °C in 200 μl ATAC resuspension buffer , pipetted up and down three times and incubated on ice for 3 min. Then 1 ml ATAC wash buffer was added, and mixed by inverting the tube three times. Nuclei were pelleted by centrifugation at 500for 10 min at 4 °C on swinging bucket rotor centrifuge with appropriate adapters for microtubes.
Upon completion, the sample was recovered by post-GEM incubation cleanup with Dynabeads MyOne SILANE and SPRIselect beads according to Chromium Next GEM Single Cell ATAC Reagent Kits v1.1 After elution from SPRIselect beads, 40 μl sample was recovered and 2 μl were used to measure the concentration using Qubit dsDNA HS Assay kit . The rest of the sample was used for P7 tagmentation by mixing the following reaction: 38 μl template DNA, 50 μl 2× TD buffer , 1 μl MeB-only loaded standard Tn5 per 10 ng template DNA and dHO up to 100 μl and incubation for 30 min at 37 °C in a thermocycler.
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