AMPLICON of Severe acute respiratory syndrome coronavirus 2: Human NJ-BioR-745-QIAseq
The NJ-BioR-745-QIAseq library was prepared using the QIAseq FX DNA Library UDI Kit (QIAGEN) in accordance with manufacturer's recommendations. Briefly, 5uL of RNA was reverse transcribed followed by amplification of cDNA targets using the QIAseq SARS-CoV-2 Primer Panel (QIAGEN) using 35 cycles of PCR and cleaned-up by magnetic bead purification. The amplicons were then partially digested, ligated to QIAseq UDI adapters, and amplified using 6 cycles of PCR. Final libraries were cleaned-up by magnetic bead purification and quantified using fluorescent-based assays including PicoGreen (Life Technologies) or Qubit Fluorometer (Invitrogen), and Fragment Analyzer (Advanced Analytics). Final libraries were sequenced on Illumina NovaSeq 6000 with 2x150bp reads targeting 0.5M read coverage.