This review focuses on the rapidly growing field of natural 238U/235U variability, largely driven by the technical advances in the measurement of U isotope ratios by mass spectrometry with increasing precision over the last decade. A thorough review on the application of the U-decay series systems within Earth sciences was published in Reviews in Mineralogy and Geochemistry (RiMG) volume 52 in 2003, and will not be discussed further within this review. Instead, this article will first focus on the basic chemical properties of U and the evolution of 238U/235U measurement techniques, before discussing the latest findings and use of this isotopic system to address questions within geochronology, cosmochemistry and Earth sciences.