Amplification and Sequencing of Norovirus Genes

Amplification and Sanger sequencing of a larger fragment of the RdRp and VP1 gene were carried out on NoV positive samples as previously described^{10 (link),29} . Briefly, one-step RT-PCR was carried out using “SuperScript III One-Step RT-PCR System kit” with Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA) following thermal profile was applied: retro-transcription at 50 °C for 60 min, initial denaturation at 94 °C for 2 min followed by 40 cycles with Taq activation at 94 °C for 15 sec, annealing at 55 °C for 30 sec, extension at 68 °C for 90 sec and final elongation at 68 °C for 5 min.
The sequences obtained were compared with those available using BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi), and nt sequences were deposited into GenBank under accession numbers: MN605609-MN605633.