Schematic of iscChIC-seq. (A) Experimental flow. (1) Bulk cells were split into the first 96-well plate after antibody-guided MNase cleavage and end repair. (2) Barcoded cells were pooled together and sorted into the second 96-well plate to introduce the i7 index. (3) Cells were pooled together again from each plate and labeled with the i5 index in PCR2. (B) Illustration of poly(dG) addition to DNA ends by TdT, oligo dC adaptor ligation by T4 DNA ligase, and PCR-mediated barcoding process. Cell barcode (red) is designed into the oligo dC P7 adaptor in which 3′ ends are blocked to prevent nontemplate tailing by TdT. After reverse crosslinking, barcoded DNA fragments could be efficiently labeled with the i7 index (purple) through annealing and PCR extension. The barcoded P5 adaptor is added to the other end of genomic DNA fragments by ligation and PCR2, which is used to amplify the library DNA for NGS sequencing.
