title: Amplicon Sequencing of rVSV/SARS2: SARS-CoV-2 Spike
design: RNA was isolated using Trizol-LS and reverse transcribed using random hexamer primers and Superscript III reverse transcriptase (Thermo Fisher Scientific, US). The cDNA was amplified using Phusion (NEB, US) and primers flanking the entire S-encoding sequence. The PCR products were gel-purified, ligated to unique i5/i7 barcoded primer combinations using the Illumina Nextera XT Index Kit v2 and KAPA HiFi HotStart ReadyMix (2X; KAPA Biosystems) and purified using AmPure Beads XP (Agencourt).