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Raman-based distributed temperature sensing (DTS) is a valuable tool for field testing and validating heat transfer models in borehole heat exchanger (BHE) and ground source heat pump (GSHP) applications. However, temperature uncertainty is rarely reported in the literature. In this paper, a new calibration method was proposed for single-ended DTS configurations, along with a method to remove fictitious temperature drifts due to ambient air variations. The methods were implemented for a distributed thermal response test (DTRT) case study in an 800 m deep coaxial BHE. The results show that the calibration method and temperature drift correction are robust and give adequate results, with a temperature uncertainty increasing non-linearly from about 0.4 K near the surface to about 1.7 K at 800 m. The temperature uncertainty is dominated by the uncertainty in the calibrated parameters for depths larger than 200 m. The paper also offers insights into thermal features observed during the DTRT, including a heat flux inversion along the borehole depth and the slow temperature homogenization under circulation.