Multifactorial profiling of epigenetic landscapes at single-cell resolution using MulTI-Tag
). In brief, nuclei were thawed and bound to washed paramagnetic concanavalin A beads and then incubated with primary antibody at 4 °C overnight in Wash Buffer with 2 mM EDTA. Bound nuclei were washed and incubated with secondary antibody for 1 hour at room temperature and then washed and incubated in Wash-300 Buffer with 1:200 loaded pA–Tn5 for 1 hour at room temperature.
. After amplification, microwell contents were re-aggregated, and libraries were purified with two rounds of cleanup with 1.3× HighPrep beads and resuspended in 20 µl of 10 mM Tris-HCl with 1 mM EDTA.Libraries were sequenced on an Illumina HiSeq instrument with paired-end 25 × 25 reads. Sequencing data were aligned to the UCSC hg19 genome build using Bowtie2 , version 2.2.5, with parameters –end-to-end–very-sensitive–no-mixed–no-discordant -q–phred33 -I 10 -X 700.
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