The motivation of this study is stimulated by a lack of knowledge about the difference of airflow characteristics between a novel air distribution method i.e., stratum ventilation (SV) and conventional air distribution methods i.e., mixing ventilation (MV) and displacement ventilation (DV). Detailed air velocity and temperature measurements were conducted in the occupied zone of a classroom with dimensions of 8.8 m (L) × 6.1 m (W) × 2.4 m (H). Turbulence intensity and power spectrum of velocity fluctuation were calculated using the measured data. Thermal comfort and cooling efficiency were also compared. The results show that in the occupied zone, the airflow characteristics among MV, DV, and SV are different. The turbulent airflow fluctuation is enhanced in this classroom with multiple thermal manikins due to thermal buoyancy and airflow mixing effect. Thermal comfort evaluations indicate that in comparison with MV and DV, a higher supply air temperature should be adopted for SV to achieve general thermal comfort with low draft risk. Comparison of the mean air temperatures in the occupied zone reveals that SV is of highest cooling efficiency, followed by DV and then MV.