Two-step RT-PCR assay workflow

A two-step RT-PCR workflow was planned and run for the detection of an RNA target, with RNA converted to cDNA before amplification. Each run included no-template and positive controls and technical replicates. Amplification curves and quality indicators were reviewed against the controls before any result was reported, together with its limitations.

Techniques used

• RT-PCR
• Assay design
• Amplification data review