GSM9848715: DVG7TD_DVG-A 2.fastq.gz; Severe acute respiratory syndrome coronavirus 2; RNA-Seq
Total RNA was extracted from infected cells using Trizol according to the manufacturer's instructions. Total RNA was extracted from DVG-B stocks using Trizol-LS according to the manufacturer's instructions. Total RNA was DNase digested and RNA was re-purified using lithium chloride. RNA-sequencing of infected A549-ACE2 cells was performed by Plasmidsaurus. For sequencing library preparation, Poly-A mRNA capture and reverse transcription for cDNA synthesis: oligo-dT primer to capture just the mRNA (contains sample barcode, UMI, and Read 1 sequences). Second strand synthesis and tagmentation: double stranded-cDNA is generated. Tagmentation creates fragments and incorporates Read 2 sequence. Illumina library amplification: adds unique dual indices (UDI = i5, i7) and P5/P7 sequences. RNA-sequencing of Total RNA from DVG-B stocks was perfromed by Genewiz. RNA quality was assessed using the RNA Pico 6000 module on an Agilent Tapestation 2100 (Agilent Technologies) prior to cDNA library preparation. 100 ng of total cDNA libraries were prepared using the Illumina TruSeq Stranded Total RNA LT kit with Ribo-Zero Gold, according to the manufacturer's instructions. Samples were run on Illumina NextSeq 500 to generate 150 bp, paired-end reads, resulting in 20-50 million reads per sample, with an average Q30 score ≥ 96.8%.