GSM6395384: COVIRS_18_22-00252_V3A; Homo sapiens; RNA-Seq
RNA extraction and genomic DNA elimination was carried out using the RiboPure RNA Purification kit for blood (Invitrogen) as per the manufacturer's instructions. Final elution into 50 uL RNase-free water. A further RNA precipitation reaction was carried out. RNA was resuspended 2.5x in 100% ethanol and 10% sodium acetate (Merck) and spun at 12,000 x g for 15 min at 4°C. Samples were washed in 75% ethanol. Pellets were air dried and resuspended in 29 uL RNase free water and total RNA yield was determined by analysis of samples using a TapeStation (Agilent, Santa Clara, USA) and Qubit (ThermoFisher Scientific, Waltham, USA). Total RNA was converted to strand-specific Illumina compatible sequencing libraries using the Nugen Universal Plus Total RNASeq library kit from Tecan (Mannedorf, Switzerland) as per the manufacturer's instructions (MO1523 v2) using 12 cycles of PCR amplification for the final libraries. An Anydeplete probe mix, targeting both human ribosomal and adult globin transcripts (HBA1, HBA2, HBB, HBD), was used to deplete these transcripts. The final library pool was converted to an MGI compatible circularized template using the MGIEasy Universal Library Conversion Kit (MGI, Shenzhen, China) before sequencing of the library pool (2 × 150 bp paired-end reads) was performed on an MGI DNBSEQ-G400.