In this study we investigated effect of the time of the day when manual measurements of soil CO2 efflux are performed on estimates of seasonal sums of released carbon from the soil. We subsampled continuous measurement of soil CO2 efflux into six sets of data in accordance to the time of the day when the measurements were taken – 0 h, 4 h, 8 h, 12 h, 16 h and 20 h. To estimate seasonal carbon flux from the soil we used continuously measured soil temperature and parameters R10 (soil CO2 efflux normalized for temperature of 10 °C) and Q10 (the proportional change in CO2 efflux caused by 10 °C increase in temperature) calculated from continuous measurements and from measurements taken at individual hours. Values of Q10 calculated from 12 h and 16 h data were lower than Q10 calculated from continuous measurements. On the contrary, Q10 at 0 h, 4 h, 8 h and 20 h were higher. Seasonal carbon flux from the soil based on 0 h, 4 h and 8 h measurements was overestimated compare to the flux calculated from continuous measurements. On the contrary, measurements at 12 h, 16 h and 20 h measurements underestimated the carbon flux. The under- or overestimation was significant for 0 h, 4 h, 8 h and 20 h data sub-sets.