The objective of this design note is to discuss and define the total uncertainty in temperature calibrations by comparison, by analysing most of the likely error sources. As a result of the proposed and developed uncertainty analysis, further reductions of uncertainties could be realized if/when better equipment becomes available. The analysis is performed as a case study using state-of-the-art calibration equipment described in the design note. This equipment is located in the authors' own secondary temperature calibration laboratory. Accreditation for this laboratory has been granted through The Dutch Council of Accreditation (RVA) for calibrations in the temperature range -55 to C. In temperature calibrations by comparison the four main groups of uncertainties are the reproducibility, uncertainty of a reference thermometer, uncertainty of a calibration bath or a furnace and uncertainty of a measuring device. Special care is taken, using a thorough evaluation procedure, to ensure that the uncertainty contribution of the calibration bath or furnace is as low as possible. This is necessary because the total uncertainty assigned to an instrument under calibration is larger than the largest individual uncertainty contribution. In temperature calibrations the largest uncertainty is most likely to be the uncertainty of the calibration bath or a furnace. Therefore this uncertainty typically represents the lowest limit for further reduction of the total uncertainty of the calibration process. The analysis performed allows optimal use of temperature calibration equipment for calibration of thermometers by comparison. In this way most practical calibration needs are satisfied in a more economical way than by using substantially more expensive fixed point calibrations.