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Single-molecule experimental design for measuring RT binding to the vRNA-tRNA complex. (a) Two different RNA oligonucleotides are annealed to the vRNA to promote formation of either an 18 bp PBS helix or 22 bp PBS helix which extends into the RNase H domain of RT.
(b) Design of mutant vRNAs with variable PBS helix lengths in complex with full-length tRNA.
