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First, viral RNAs were extracted directly from clinical samples with the QIAamp Viral RNA Mini Kit, and then used to synthesise cDNA with the SuperScript III Reverse Transcriptase (ThermoFisher, Waltham, MA, USA) and N6 random primers, followed by second-strand synthesis with DNA Polymerase I, Large (Klenow) Fragment (ThermoFisher). Viral cDNA libraries were prepared with use of the Nextera XT Library Prep Kit (Illumina, San Diego, CA, USA), then purified with Agencourt AMPure XP beads (Beckman Coulter, Brea, CA, USA), followed by quantification with an Invitrogen Qubit 2.0 Fluorometer. |
OTHER |
METATRANSCRIPTOMIC |
RT-PCR |
PAIRED
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