Qiaamp 96 viral rna kit

Viral RNA was extracted from 180 µl of each saliva sample using the QIAamp 96 Viral RNA Kit with the QIAcube HT (Qiagen) using the following settings with a filter plate: the lysed sample was pre-mixed 8 times before subjecting to vacuum for 5 min at 25 kPa and vacuum for 3 min at 70 kPa. Following 3 washes using the same vacuum conditions above, the samples were eluted in 75 µl AVE buffer followed by a final vacuum for 6 min at 60 kPa. Next, nine microliters of RNA were used for cDNA synthesis and library preparation using the Illumina COVIDSeq Test kit (Illumina) and Mosquito HV Genomics Liquid Handler (SPT Labtech Inc.). The size and purity of the library were determined using the 4200 TapeStation System (Agilent) and the Qubit dsDNA HS Assay Kit (Life Technologies), according to the manufacturer's instructions. Constructed libraries were pooled and sequenced using the NovaSeq 6000 Sequencing System SP Reagent Kit and the NovaSeq Xp 2-Lane Kit. Illumina's DRAGEN pipeline was used to derive sample consensus sequences, which were filtered based on a minimum of 70% coverage of the genome.

Viral RNA was extracted from 180 μl of each saliva sample using the QIAamp 96 Viral RNA Kit with the QIAcube HT (Qiagen) using the following settings with a filter plate: the lysed sample was premixed eight times before subjecting to vacuum for 5 min at 25 kP and vacuum for 3 min at 70 kPa. Following three washes using the same vacuum conditions above, the samples were eluted in 100 μl AVE buffer followed by a final vacuum for 6 min at 60 kPa. Nine microliters of RNA was used for cDNA synthesis and library preparation using the COVIDSeq Test kit (Illumina) and Mosquito HV Genomics Liquid Handler (SPT Labtech Inc.). The size and purity of the library were determined using the 4200 TapeStation System (Agilent) and the Qubit dsDNA HS Assay Kit (Life Technologies) according to the manufacturer's instructions. Constructed libraries were pooled and sequenced using the NovaSeq. 6000 Sequencing System SP Reagent Kit and the NovaSeq Xp 2‐Lane Kit. Illumina's DRAGEN pipeline was used to derive sample consensus sequences, which were filtered based on a minimum of 70% coverage of the genome.