For over 25 years, real-time quantitative polymerase chain reaction (qPCR), also known as quantitative PCR or real-time PCR (RT-PCR or RT-qPCR), has offered a comparatively cheap and rapid means of quantifying gene abundances in a diverse range of samples (VanGuilder et al., 2008) including environmental samples such as soils. This has allowed unprecedented insight into important microbial metrics such as the abundance of different bacterial taxa (Fierer et al., 2005, Zhang et al., 2017) and the abundance of process-specific functional genes, enabling their associations with functional measures, e.g. volatile organic compounds or greenhouse gas emissions (Shahsavari et al., 2016; Giles et al., 2017). In addition, it has allowed the creation of diagnostic and monitoring tools for both soil prokaryotes and eukaryotes (Jiménez-Fernández et al., 2010). This has revolutionised the ability of microbial ecologists to both examine changes in soil communities, as well as identify and quantify populations of beneficial and pathogenic organisms, thereby generating a better understanding of the role microbial soil communities play in supporting soil function and broader ecosystem services. Using qPCR to investigate soil communities, however, has a wide range of advantages, disadvantages, and potential limitations that need to be accounted for when designing, implementing, and interpreting qPCR-based studies. The continued improvement of qPCR approaches, together with their integration into the growing range of other molecular methods targeted at exploring soil communities, means that qPCR remains one of our most powerful tools for studying soil microbial communities and dissecting their various roles in soil health.