Soil respiration measurements are commonly used as soil health indicators. Several ex situ soil respiration methods exist, but comparative performances between them have rarely been analyzed. Specifically, there is a lack of comparisons between intact microcosms and destructive methods. The objective of this study was to analyze and compare three different ex situ soil respiration methodologies: minimally disturbed microcosms using fresh soil, dried–sieved 24 h burst test, and dried–sieved 10-day incubation. We hypothesized that (i) the respiration rates for the three methods are correlated to each other; (ii) the respiration rates are strongly correlated with soil physico-chemical parameters; (iii) disturbance caused by drying and sieving reduces regression coefficients compared with microcosms; and (iv) drying and sieving soil produces larger respiration rates. Soil was collected in the Province of New Brunswick, Canada. Total carbon and nitrogen (C:N), pH, aggregate stability, total dissolved C and N, NO₃ and NH₄, texture, and labile C were determined prior to incubations. Our results showed that the three methods had CO₂ efflux in similar ranges. However, all the methods had low to no significant correlations between soil physico-chemical parameters and respiration. Total dissolved N had the strongest correlation with CO₂ efflux. The results of the microcosm method significantly correlated with the results for 24 h burst test but not with the 10-day incubation method. We conclude that drying and sieving soil prior to performing ex situ soil heterotrophic respiration measurements using the 24 h burst tests can produce cautiously reliable results. Despite the disturbance, results from the 24 h burst tests are comparable with the results of the microcosm method.