title: RNA-Seq of HCV: WT, control
design: Amplicon sequencing of four overlapping NS5A/NS5B cDNA fragments of HCV; cDNA synthesized with SuperScript IV; PCR with Phusion High-Fidelity DNA Polymerase (98C 30s; 35 cycles of 98C 10s, 60C 30s, 72C 30s; final 72C 2min); amplicons gel-purified and quantified by Qubit; libraries prepared with NEBNext Ultra II FS DNA (NEB E7805); NextSeq 550 run, 2125 bp; pooled 5 nM, diluted to 4 nM; 1.6 pM loaded with 1% PhiX.