Mutations in the JAK2 gene are prevalent in the human myeloid malignancies, being present in virtually all cases of polycythemia vera, and a significant proportion of patients with other myeloproliferative disorders. Various methods for the detection of acquired mutations in this gene are available depending on the need for sensitivity, quantification, or the ability to detect many different mutations. We summarize the various methods published and discuss their relative merits for each application. Two commonly used methods, quantitative real-time PCR (QPCR) for the detection of the JAK2 V617F mutation and high resolution melt-curve analysis (HRM) for the detection of multiple mutations within JAK2 exon 12, demonstrate the utility of each method and their limitations. The choice of methodology is dependent on the application; therefore there is no gold standard for detecting mutations in this gene.