ADT Data from CITE-seq of BNT162b2 mRNA-Vaccinated Donor PBMCs
RNA library construction was performed according to the 10x scRNA-seq protocol, while the ADT (oligo-tagged antibodies of cell surface proteins) and HTO (cell hashing tags) library constructions were conducted following the CITE-seq protocol (https://citeseq.files.wordpress.com/2019/02/cite-seq_and_hashing_protocol_190213.pdf). During cDNA amplification (Step 2.2a), 0.2 μM of ADT additive primer (5′CCTTGGCACCCGAGAATTCC) and 0.1 μM of HTO additive primer (5′GTGACTGGAGTTCAGACGTGTGCTC) were added to the reaction mixture to enrich antibody tags. During cDNA cleanup (Step 2.3), supernatant containing the antibody tags was saved and further purified with 2x SPRI. The eluate was split into two tubes for ADT and HTO libraries. After cDNA cleanup, additional PCR reactions generated ADT and cell hashing libraries. These reactions were set up with KAPA Hifi Master Mix with the following primers: 10 μM 10x Genomics SI-PCR primer (5′AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTC), and 10 μM Illumina TruSeq DNA D7xx primer (5′CAAGCAGAAGACGGCATACGAGATxxxxxxxxGTGACTGGAGTTCAGACGTGTGC) for HTO library. 10 μM 10x Genomics SI-PCR primer, and 10 μM TruSeq Small RNA RPIx primer (5′CAAGCAGAAGACGGCATACGAGxxxxxxxxGTGACTGGAGTTCCTTGGCACCCGAGAATTCCA) for ADT library. The PCR products were purified with 1.6x SPRI.