Lc isolink interface

Soil microbial total DNA-C concentration was used as a proxy for biomass C; DNA extraction was carried out on a soil aliquot of 0.25 g using PowerSoil-htp 96-well soil DNA isolation kit following the manufacturer's instructions (MO BIO Laboratories, UK). Another set of identical DNA extraction was performed following addition of 25 µL of DO¹³C solution and overnight (16 h) incubation in the dark at field moisture capacity. Both extracts were analyzed in the size-exclusion chromatographic (SEC) mode on a liquid chromatography isotope ratio mass spectrometer LC-IRMS (HPLC system coupled to a Delta + XP IRMS through an LC IsoLink interface; Thermo Fisher Scientific, Germany)^{46 (link)}. This allowed us to obtain the DNA-C content and the proportion of DO¹³C in microbial DNA from soils with and without substrate addition. Microbial CUE was estimated as DNA-¹³C/(DNA-¹³C + ∑CO₂-¹³C), where ∑CO₂-¹³C is the cumulative DO¹³C lost during respiration. Microbial turnover rate (synonymously referred to as growth rate) was calculated as DNA-¹³C/DNA-C.

Frozen soil (1 g) was placed in a 10 mL glass vial, thawed and incubated for approximately 24 h in the dark at room temperature. Microbially respired ¹³CO₂ collected in the headspace was measured using a gas chromatography isotope ratio mass spectrometer (GC-IRMS, Delta⁺ XL, Thermo Fisher Scientific, Germany) coupled to a PAL-autosampler (CTC Analytics) with general purpose (GP) interface (Thermo Fisher Scientific, Germany). Microbial biomass from soil was obtained using a slightly modified version of the CFE method (Vance et al., 1987 (link); Malik et al., 2013 (link)). Soil (7 g wet weight) was fumigated with chloroform gas and organic carbon was extracted from both fumigated and non-fumigated soils. Soil extracts were analyzed in the bulk (μEA) mode on an HPLC-IRMS (HPLC system coupled to a Delta⁺ XP IRMS through an LC IsoLink interface; Thermo Fisher Scientific, Germany).