AIRR-Seq of Homo sapiens: Blood samples from BNT162b2 vaccinees
Total RNA was used to synthesize cDNA, using the Superscript IV First-Strand Synthesis System (Invitrogen), with primers targeting the immunoglobulin heavy chain CH1 domain of each isotype according to the manufacturers protocol. SPRI beads (AmpureXP, Beckman Coulter, Brea, CA, USA) were used to purify cDNA, which was subjected to second-strand synthesis using V gene-specific primers and KAPA Biosystems (KAPA HiFi HotStart, Roche, Basel, Switzerland). dsDNA was purified using SPRI beads and VH genes were amplified. Sequencing libraries were quantified with a 4200 TapeStation System (Agilent Technologies), using a D1000 ScreenTape Assay.