•Inexpensive and reliable approach for characterisation of fractured aquifers.•Combines uniform and point injection dilution tests on open sections of boreholes.•Proposed workflow to identify nature and depth distribution of flowing fractures.•Fracture porosity and permeability distribution determined for specific conditions.•Groundwater velocities determined were validated against borehole-to-borehole tests. Fractured carbonate aquifers derive their transmissivity essentially from a well-developed network of solutionally-enhanced fractures and conduits that can lead to high groundwater velocities and high vulnerability to contamination of water quality. Characterisation of the variation of hydraulic properties with depth is important for delineating source protection areas, characterising contaminant fate and transport, determination of the effectiveness of aquifer remediation, and parameter estimation for models. In this work, ambient open borehole uniform and point injection dilution tests were conducted on observation boreholes in the unconfined Cretaceous Chalk aquifer of East Yorkshire, UK, and interpreted in conjunction with other data via the implementation of a new work flow. This resulted in the characterisation of flow in these boreholes and the inference of properties such as groundwater flow patterns and velocities in the surrounding aquifer formation. Our workflow allowed sections of open boreholes showing horizontal versus vertical flow to be distinguished, and the magnitude of such flows and exchanges with the aquifer to be determined. Flow within boreholes were then used to characterise: i) presence and direction of vertical hydraulic gradients; ii) nature and depth distribution of flowing features; iii) depth interval porosity and permeability estimation of the flowing features from overall borehole transmissivity and geophysical image or caliper logs; iv) groundwater velocity estimation in the surrounding aquifer. Discrete flowing features were distributed across the range of depths sampled by the observation boreholes (typically up to 45–60 mbgl), but the majority were located in the zone of water table fluctuation marked by solutionally enlarged flow features. Quantitative interpretation of both uniform injection (tracer distributed throughout the open borehole section) and point injection (slug of tracer introduced at targeted depth) yielded vertical velocities within the borehole water column in broad agreement with those measured by flow logging. Depth specific fracture kinematic porosities inferred from the ambient dilution data combined with long-interval pump test and geophysical log data ranged between 3.7 × 10−4–4.1 × 10−3 with an average of 2.1 × 10−3; these values were in excellent agreement with those from other methods applied to the same aquifer such as larger scale pumping tests. A new approach to estimation of groundwater velocities from the dilution test data using externally measured hydraulic gradients gave inferred horizontal groundwater velocities ranging between 60 and 850 m/day, in full agreement with those from previously conducted borehole-to-borehole tracer tests. These results confirm that the studied aquifer is karstic, with rapid preferential pathways which have implication for flow and transport modelling, and pollution vulnerability. Our study results indicate that ambient single-borehole dilution approaches can provide an inexpensive and reliable approach for the characterisation of fractured and karstic aquifers.