Colorectal cancer (CRC), the second most common cause of cancer-related mortality worldwide, is preventable with effective screening and removal of precursor lesions. Yet, screening efforts have been hampered by low participation rates and by performance limitations of the screening tools themselves. Stool DNA testing has emerged as a biologically rational and user-friendly strategy for the non-invasive detection of both CRC and critical precursor lesions. Unlike most conventional screening tools, stool DNA testing detects proximal and distal colorectal neoplasms equally well. Several key technical advances have led to increasingly accurate approaches for stool DNA testing including use of a DNA preservative buffer with stool collection, efficient target capture and amplification methods, broadly informative marker panels, and automated assay components. Based on recent studies, advanced multi-marker stool DNA tests including methylated markers, mutation markers and an assessment of faecal haemoglobin have been shown to detect CRC at sensitivities of 85% and higher and adenomas >1 cm at 60% and higher in a case-control environment. If the high accuracy of multi-marker stool tests is corroborated in multicentre screening studies on average-risk persons currently underway, then these stool tests could influence our CRC screening paradigm.This review discusses the biological basis, key technical advances, and recent clinical performance validation of stool DNA testing.