FAQ: Should I include a No-RT control reaction?

No-RT control reactions are useful for determining issues that may arise from the amplification of gDNA that may be present in a sample. In general, the Luna Cell Ready One-Step RT-qPCR Kit offers efficient genomic DNA removal (>95%). For example, with an abundant transcript such as β-actin, genomic DNA is detected approximately 10 cycles later than from RNA detection after DNA removal. However, many RNA transcripts are present at low abundance. For these transcripts, it is recommended to perform a No-RT control reaction, particularly if primer sets do not span exon-exon junctions. In addition, no template control (NTC) reactions should be included to demonstrate that positive reactions are meaningful.