Sup. Table 1. Primers Primers Sequences (5` to 3`) Region
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Sup. Table 1. Primers Primers Sequences (5' to 3') Region✝ Nucleotides✝ Whole reverse GGGAGCAGGTAGATGCCTACCC NS5B 9331-9352 NS3-F1 ACTTCTAGGACCGGCCGATA NS2 3359-3378 NS3-R1 GCTCGTCACACTTCTTCTTG NS3 4517-4536 NS3-F2 CCAATCGCCCAGATGTACAC NS3 3615-3634 NS3-R2 GTCTGTGAAGACCGGAGACC NS3 3946-3965 NS5A-F1 TACTCCCTGCCATCCTCTCTCCTG NS4B 5974-5997 NS5A-R1 CTCCTTGAGCACGTCCCGGT NS5B 7777-7796 NS5A-F2 CTGTGCAGTGGATGAACCGG NS4B 6064-6083 NS5A-R2 AGGAGAGGTTTGCACACTGG NS5A 6736-6751 NS5A-F3 GTTGCTGAAGAGGCTTCATCAGT NS4B 6194-6216 NS5A-R3 CACAGCGCCCTGGAATAGTT NS5A 6558-6577 NS5B-F1 TCAGCGACGGGTCTTGGTCTAC NS5A 7528-7549 NS5B-F2 AGCGGGTCGAGTTCCTGGTG NS5B 8182-8201 NS5B-F3 GGGTCGAGTTCCTGGTGAAT NS5B 8185-8204 NS5B-R1 AGATAACGACAAGGTCGTCTCC NS5B 8535-8556 ✝ HCV-J (accession No. D90208) was used as a reference. 1 Sup. Table 2. PCR conditions NS3 NS5A NS5B second PCR for second PCR for first PCR population sequencing deep sequencing primer pairs primer pairs primer pairs NS3-F1/NS3-R1 NS3-F1/NS3-R2 NS3-F2/NS3-R2 temperature time cycle 94 2min ×1 98 10s 56 30s 68 50s 68 5min ×35 ×1 temperature time cycle 94 2min 94 3min 98 10s 94 10s 56 30s 68 40s 68 5min ×35 temperature time 58 30s second PCR for first PCR population sequencing deep sequencing primer pairs primer pairs primer pairs NS5A-F1/NS5A-R1 NS5A-F2/NS5A-R2 NS5A-F3/NS5A-R3 time cycle 94 2min ×1 98 10s 68 70s 68 5min ×35 ×1 temperature time cycle 94 2min 94 3min 98 10s 94 10s 60 30s 68 40s 68 5min ×35 temperature time 58 30s second PCR for first PCR population sequencing deep sequencing primer pairs primer pairs primer pairs NS5B-F1/Whole reverse NS5B-F2/Whole reverse NS5B-F3/NS5B-R1 temperature time cycle temperature time temperature 94 2min ×1 94 2min 98 10s 98 10s 68 70s 68 45s 68 5min 68 5min ×1 2 ×35 cycle ×1 ×35 68 90s second PCR for ×35 ×1 68 90s second PCR for temperature cycle cycle time 94 3min ×35 cycle ×1 94 10s 58 30s 68 90s ×35 Sup. Materials and Methods Generation of humanized NOG mice NOG mice expressing a thymidine kinase transgene (TK-NOG)-based human hepatocyte chimeric mice were generated as previously reported [10]. Briefly, eight-week-old TK-NOG mice were injected intraperitoneally with 10 mg/kg of ganciclovir (Mitsubishi Tanabe Pharma Corporation, Osaka Japan). After one week, mice were transplanted with 1×106 of human hepatocytes. Blood of transplanted mice were collected from external jugular vein every 2-4 weeks and serum levels of human albumin, which correlated with the human hepatocyte chimeric rate [10], were measured by ELISA using Human Albumin ELISA Quantitation Set (Bethyl Laboratories Inc., TX). 3
