Influenza Virus High-Throughput Influenza Sequencing Pipeline
- Sample collection and amplification by cell culture
- Extraction of virus genomic RNA from cell culture supernatants (48 samples at a time)
- RNA shipped to TIGR and processed in 96-well plate format
- Primer design
- Amplification by RT-PCR of overlapping regions of each virus segment (1 virus isolate per 96-well plate; 4 plates per thermocycler)
- Sequencing of PCR amplicons using M13-universal primers in 384-well plate formats (4 virus isolates sequenced simultaneously)
- Trimming of sequences, assembly of segments, editing of nucleotides
- Quality assurance and submission to NCBI
RSS