Fig. 1. ApharSeq overview.

(A) Barcoded and uniquely identifiable RT primers are hybridized to samples in transport/lysis buffer, and homemade paramagnetic polyT beads are used for a quick wash and buffer replacement step. (B) Beads are pooled and RNA undergoes an RT/PCR with prehybridized target-specific barcoded primers to generate a sequencing library. (C) Libraries are sequenced and analyzed, and PCR duplicates are collapsed to molecular counts for detection and further analysis (e.g., major variant calling by sequence analysis and contact tracing by minor variations).