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Amplification and sequencing of HCV NS5B Viral RNA was extracted from plasma with the QIAmp Viral RNA Mini kit (Qiagen, Hilden, Germany) and a 372 bp region of NS5B gene corresponding to the Okamoto region was amplified using Titan One Tube RT-PCR System (Roche Diagnostics) with the primers HCV_FW1 [5-CCCGCTGYTTTGACTCVACNGT-3, location in GT1a isolate H77 (GenBank accession number AF009606), 82648285] and HCV_RV1 (5-CCTRGTCATAGCCTCCGTGAA-3, location 86368616)

Nicol requests a novelty number called My Canary Has Circles Underneath His Eyes and says he hopes to record it with Yoko Ono and John Lennon

(R Foundation for Statistical Computing, 2020)

doi: 10.3892/etm.2015.2964 74