Inter and intra-host diversity of RSV in hematopoietic stem cell transplant adults with normal and delayed viral clearance
RNA was extracted from patient [stool/nasal samples or infected human intestinal organoids] and RNA extracts were converted to cDNA using NEBNext RNA First Strand Synthesis Module and NEBNext Ultra II Directional RNA Second Strand Synthesis Module. Illumina unique dual barcodes adapters were ligated onto purified cDNA and libraries were amplified using the Kapa HiFi HotStart Library Amplification kit. Pooled cDNA libraries were hybridized with biotin-labeled probes from the [HuNoV OR RSV] Panel (Twist Biosciences, Inc) at 70C for 16?hours. The post-capture pool was further PCR amplified for 12-20 cycles and final libraries were sequenced on Illumina NovaSeq 6000 S4 flow cell, to generate 2150 bp paired-end reads.