ADT Data from ASAP-seq of BNT162b2 mRNA-Vaccinated Donor PBMCs
ASAP-seq was conducted as previously described with minor modifications. After staining with cell surface antibodies, cells were fixed in 0.1% formaldehyde for 5 minutes at room temperature. After washing, the cell pellet was resuspended in 100 µL of lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% Nonidet-P40 substitute (IGEPAL), 1% BSA) and kept on ice for 5 mins. The permeabilized cells were then resuspended with 1× Diluted Nuclei Buffer (10x Genomics) to a concentration of around 5000 cells/µL. 10 µL transposition mix (3µl 10x ATAC Buffer B and 7µl 10x ATAC Enzyme) was mixed with 5 µL sample and incubated for one hour at 37 °C. 0.5 μM bridge oligo A (TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGNNNNNNNNNVTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT/3InvdT/) was added to the barcoding mix for proper amplification of antibody tags. The GEM incubation was performed with the following PCR program: 40 °C for 5 min, 72 °C for 5 min, 98 °C for 30 s; 12 cycles of 98 °C for 10 s, 59 °C for 30 s and 72 °C for 1 min; ending with hold at 15 °C. Chromatin accessibility (ATAC), hashtags (HTOs) and oligo-tagged antibodies of cell surface proteins (ADTs) libraries were constructed by following the ASAP-seq protocol (https://citeseq.files.wordpress.com/2020/09/asap_protocol_20200908.pdf).