Creating cDNA Libraries using Twist Library Preparation Kit for ssRNA Virus Detection
Most viral pathogens are single-stranded RNA including SARS-CoV-2, the virus behind the 2019–2020 Coronavirus pandemic.
This protocol describes how to convert RNA into cDNA libraries for downstream use in NGS Target Enrichment. The protocol begins with creating cDNA from extracted/purified RNA using commercially available reagents.
Then cDNA is converted to Illumina TruSeq-compatible libraries using the Twist Library Preparation Kit using Enzymatic Fragmentation and Unique Dual Indices (UDI) system. Synthetic viral RNA controls, such as the Twist synthetic SARS-CoV-2 RNA controls, can be spiked into reference RNA samples to serve as a positive control. The cDNA libraries are compatible with Twist Target Enrichment workflows.