WizPure™ qRT-PCR Master (EVA) is an optimized ready-to-use solution for one-step quantitative RT-PCR assays, incorporating EvaGreen dye. It comprises all the components necessary to perform qRT-PCR: WizScript™ III RTase, antibody-mediated hot-start Taq DNA Polymerase, dNTPs, MgCl2, EvaGreen dye, enhancer, and stabilizer. The user simply needs to add water, a template, and primers. An enhanced buffer allows for RT reaction temperatures up to 50°C. This can improve the detection of more difficult targets as higher RT temperatures reduce nonspecific priming and facilitate the melting of RNA secondary structures. The kit includes the components necessary for performing RT-PCR amplification and has been successfully used to DNA amplify and detect a variety of RNA targets. The kit provides a one-step, simple, robust, inexpensive assay for the detection and quantitative analysis of gene expression directly from cells or RNA with an intercalator format.