TABLE 1.
Full-genome sequencing of in vitro-transcribed RNA (2aRlucNeo)
| No. of input RNA molecules per reaction | Input RNA amt per reaction | Total no. of MiSeq reads | % reads aligned to HCV (2aRlucNeo reference sequence) | Avg coverage (no. of reads per position) | % (no. total no) for: |
|||||
|---|---|---|---|---|---|---|---|---|---|---|
| Assembled 5′ UTR length | Sequence identity of assembled 5′ UTR to reference | Sequence identity CDSa | Assembled 3′ UTR length | Sequence identity of assembled 3′ UTR to reference | S282T 10%b | |||||
| 108 | 500 pg | 351,921 | 88.7 | 9,404 | 91 (328/361) | 100 (328/328) | 100 (8,474/8,474) | 58 (136/234) | 100 (136/136) | 10.6 (688/6,464) |
| 107 | 50 pg | 292,867 | 93.8 | 7,880 | 90 (326/361) | 100 (326/326) | 100 (8,474/8,474) | 58 (136/234) | 100 (136/136) | 9.7 (874/9,036) |
| 106 | 5 pg | 1,006,481 | 93.1 | 27,462 | 91 (328/361) | 100 (328/328) | 100 (8,474/8,474) | 79 (186/234) | 100 (186/186) | 9.7 (1,024/10,552) |
| 105 | 500 fg | 283,822 | 71.8 | 6,844 | 91 (328/361) | 100 (328/328) | 100 (8,474/8,474) | 22 (52/234) | 100 (52/52) | 11 (1,144/10,364) |
| 104 | 50 fg | 457,311 | 29.2 | 4,419 | 90 (326/361) | 100 (326/326) | 100 (8,474/8,474) | 21 (48/234) | 100 (48/48) | 14.6 (786/5,392) |
| 103 | 5 fg | 1,531,207 | 4.2 | 2,091 | 84 (302/361) | 100 (302/302) | 99.98 (8,146/8,148) | 19 (44/234) | 100 (44/44) | 5.9 (16/272) |
| NTCc | 0 | 1,811,726 | 0.03 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
a
Identity was calculated as the number of exact matches to the 8,474-nucleotide-long coding sequences (CDS) of 2aRlucNeo, excluding the 5′ UTR and 3′ UTR.
b
Percent hits and hits per total number of reads of the drug-resistant NS5B mutation S282T, which was added at 10% in all dilutions.
c
NTC, no-template control.