As its name implies, single-step RT-PCR combines the synthesis of complementary DNA (cDNA) of the first strand (reverse transcript, RT) and subsequent PCR in a single reaction tube. This reaction configuration simplifies the work, reduces variations and minimizes possible contamination. One-step RT-PCR makes it easier to process a large number of samples, allowing for high-throughput applications. However, one-step RT-PCR uses specific gene primers for amplification, limiting the analysis to a few genes per RNA sample. Since the reaction is a compromise between reverse transcription and amplification conditions, one-step RT-PCR may be less sensitive and less effective in some cases. However, the use of a gene-specific primer in RT-PCR can help maximize the yield of the target cDNA and minimize amplification.