title: GSM7745103: miRNA-Seq - A549ACE2-Spike-1; Homo sapiens; miRNA-Seq
library_construction_protocol: Cells were lysed using Trizol reagent (Thermo Fisher) and RNA was extracted by phase separation. Total RNA was quantified by nanodrop (DeNovix DS-11) and TapeStation 4200 (RNA Screen Tape; Agilent Technologies, Inc) or 2100 Bioanalyzer (Total RNA Nano; Agilent Technologies, Inc). A total of 100 ng total RNA was used for library preparation via the QIAseq miRNA Library Kit (#331502, Qiagen) according to manufacturer's recommendations. Briefly, QIAseq miRNA 3' and 5' adapters were ligated to mature miRNAs. The ligated miRNAs were then reverse transcribed to cDNA using a reverse transcription (RT) primer with a Unique Molecular Index (UMI). Libraries underwent cleanup and amplification (16 PCR cycles) using a forward primer that has a unique index and the reverse primer has another unique index for dual indexing (QIAseq miRNA 96 Index Kit IL UDI, #331645, Qiagen). Successful library production, quality control, and quantification was performed using Tapestation (High Sensitivity D1000 ScreenTape). Libraries were pooled together and subjected to sequencing via a NovaSeq 6000.