In this paper, we report on experimental results to measure the total temperature of nitrogen gas at the inlet and outlet of microtubes with constant wall temperature and to quantitatively determine the heat transfer rates. Experiments were conducted with nitrogen gas flowing in a stainless steel microtube with a diameter of 524 μm and a copper microtube with a diameter of 537 μm. The temperature differences between the inlet and the wall were maintained at 3, 5, and 10 K by circulating water around the inlet and the wall. The stagnation pressures were also controlled so that the flow, with atmospheric back pressure, could reach Reynolds numbers as high as 26,000. To measure the total temperature, a polystyrene tube with a thermally insulated exterior wall containing six plastic baffles was attached to the outlet. Heat transfer rates were obtained from the gas enthalpy difference by using the pressures and the total temperatures measured at the inlet and outlet. Heat transfer rates were also compared with those obtained from the ideal gas enthalpy using the measured total temperatures and from the Nusselt number of incompressible flows. It was found that the measured total temperature at the microtube outlet was higher than the wall temperature. Also, the heat transfer rates calculated from the total temperature difference were higher than the values obtained from the incompressible flow theory.