Actual and exchangeable pH was measured using deionized water and 0.1 M solution of KCl as extracting solutions, respectively (ISO 10390: Soil quality – Determination of pH. International Organization for Standardization, ISO 2000). Total C and N contents were determined by methods of Ehrenberger and Gorbach (1973 ) using CHN catalyst (Carlo Erba NC 2500), total P was measured according to the method of Olsen and Sommers (1982 ). Available P was measured in filtrate of 5 g of air dried soil with 50 ml of 0.5 M K2SO4 solution by flow injection analysis with spectrophotometric detection using the instrument QuikChem FIA + 8000 Series (Ammerman 2001 ; Egan 2001 ). Concentrations of available Ca2+ and K+ were measured using atomic emission spectrometry method and available Mg2+ using atomic absorption spectrometry according to methods of Moore and Chapman (1986 ) and Dědina (1987 ), with solution of 1 M ammonium acetate as the extractant. All analyses were performed by the Analytical Laboratory of Institute of Botany of the Czech Academy of Sciences in Průhonice.
Nc2500
Manufactured by Carlo Erba
Sourced in Italy, United Kingdom
The NC2500 is a laboratory instrument designed for the determination of nitrogen and protein content in a wide range of sample types. It utilizes the Dumas combustion method to perform accurate and reliable analysis.
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Lab products found in correlation
8 protocols using nc2500
1
Soil Chemical Analysis Protocol
2
Microbial Biomass Carbon Analysis
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For microbial biomass C (MBC) analysis, subsamples from each of the six microcosms were randomly paired (e.g., jars 1&3, 2&4, and 5&6) and combined to form triplicate samples. MBC was measured following a standard fumigation-extraction method72 (link). Briefly, one subsample (1.33 g dry wt. equivalent) was immediately extracted by agitating for one hour in K2SO4 (0.05 M, 10 mL), while the other subsample was fumigated for 24 hours with ethanol-free chloroform (0.4 mL) prior to extraction with K2SO4. The extracts were passed through filter paper (0.45 μm, Advantec) and dissolved organic C was measured by combustion catalytic oxidation (TOC-VCPN TOC analyzer Shimadzu, Japan). A portion of the extract was freeze-dried, and total C and δ13C of the freeze-dried residue were measured using isotope ratio mass spectrometry (Delta V, Thermo Scientific, Germany) coupled to an elemental analyzer (NC2500, Carlo Erba, Italy).
3
Stable Isotope Analysis of Sediments
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Stable isotopes of nitrogen (d 15 N) and carbon (d 13 C) were analyzed using a Thermoquest (Finnigan MAT) Delta Plus XL isotope ratio mass spectrometer (IRMS) coupled with a Carlo Erba NC2500 elemental analyzer following procedures Cladoceran Response to Fish Stocking and Nutrients of Savage and others (2010) . Analysis was performed on freeze-dried sediments, and values were calibrated against the international standards of atmospheric N 2 and Vienna Pee Dee Belemnite (VPBD) for d 15 N and d 13 C, respectively. Isotopic data are presented as per mille (&). Sedimentary C/N is presented as mass ratios.
4
Soil Biogeochemical Analysis Protocol
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Actual and exchangeable pH was measured using deionized water and 0.1M solution of KCl as extracting solutions, respectively (ISO 10390: Soil quality -Determination of pH. International Organization for Standardization, ISO 2000) . Total C and N contents were determined by methods of Ehrenberger and Gorbach (1973) using CHN catalyst (Carlo Erba NC 2500), total P was measured according to the method of Olsen and Sommers (1982) . Available P was measured in ltrate of 5 g of soil with 50 ml of 0.5M K 2 SO 4 solution by ow injection analysis with spectrophotometric detection using the instrument QuikChem FIA + 8000 Series (Ammerman 2001; Egan 2001 ). Concentrations of available Ca 2+ and K + were measured using atomic emission spectrometry method and available Mg 2+ using atomic absorption spectrometry according to methods of Moore and Chapman (1986) and Dědina (1987) , with solution of 1M ammonium acetate as the extractant. All analyses were performed by the Analytical Laboratory of Institute of Botany of the Czech Academy of Sciences in Průhonice.
5
Quantifying Microbial Nutrient Uptake via IRMS
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Total
NH3–N and CO2–C uptake were determined
using isotope ratio mass spectrometry (IRMS), based on total C and
N stoichiometry, AT % 13C/12C and AT % 15N/14N ratios, and changes in sample mass before
and after gas exposure. Subsamples from each experimental treatment
were ground and weighed into tin capsules. Total 12,13C
and 14,15N of samples were measured by combustion on an
isotope ratio mass spectrometer (Thermo Finnigan MAT Delta Plus, Thermo
Electron Corporation, Waltham, MA) coupled to an elemental analyzer
(NC2500, Carlo Erba, Egelsbach, Germany). To assure complete combustion,
less than 0.5 mg of the sample was mixed with the 3-fold greater weight
of vanadium pentoxide (Sigma-Aldrich, St. Louis, MO).
Nitrogen
and C uptake were calculated according toeq 4 (shown for N), relying on the 15,14N and 13,12C AT % of samples before and after gas exposure
and the AT % of gas cylinders.
NH3–N and CO2–C uptake were determined
using isotope ratio mass spectrometry (IRMS), based on total C and
N stoichiometry, AT % 13C/12C and AT % 15N/14N ratios, and changes in sample mass before
and after gas exposure. Subsamples from each experimental treatment
were ground and weighed into tin capsules. Total 12,13C
and 14,15N of samples were measured by combustion on an
isotope ratio mass spectrometer (Thermo Finnigan MAT Delta Plus, Thermo
Electron Corporation, Waltham, MA) coupled to an elemental analyzer
(NC2500, Carlo Erba, Egelsbach, Germany). To assure complete combustion,
less than 0.5 mg of the sample was mixed with the 3-fold greater weight
of vanadium pentoxide (Sigma-Aldrich, St. Louis, MO).
Nitrogen
and C uptake were calculated according to
and the AT % of gas cylinders.
6
Elemental Analysis of Plant Biomass
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In the first harvest, at the end of the first year of the experiment, half of the trees (n = 20 per species and treatment), all tansy plants, and all shed foliage were collected. The remaining trees were then transferred to the WSL horticultural facility, where they were left for one year in ambient field conditions, until a second harvest at the end of the second year of the experiment. At each harvest, the collected material was divided into soil (tree pots only), root (cleaned coarse and fine roots < 2 mm diameter), wood (tree stems and twigs), and foliage (including the herbaceous shoots of tansy plants) fractions, and the CaCl2 pH of the soils was determined. All plant material was oven-dried at 65 °C until a constant weight was reached, and dry mass was assessed. The material within each fraction was then homogenised, subsampled for elemental analysis, and ground to a fine powder (Retsch MM2000 zirkonoxid-bowl ultra-centrifuge mill, Retsch GmbH, Hann, Germany). It was then digested in a high-pressure microwave system (240 °C, 12 MPa; ultraClave, Milestone, Sorisole, Italy) and analysed in duplicate (spread < 10%) using a gas chromatograph (NC-2500, Carlo Erba-Instruments, Wigan, UK) for C and N and by ICP-OES (Optima 7300 DV, PerkinElmer Inc., Waltham, MA, USA) for Cd, Cu, Pb, Zn, and the macronutrients Ca, K, Mg, and P according to ISO 17025.
7
Quantifying Lysozyme Incorporation in Silica Composites
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To cross-correlate the in situ SAXS data, the silica–lysozyme suspensions were dried in an oven at 40 °C for ca. 48 h. The resulting powders were washed five times with MilliQ water to remove excess lysozyme and salts followed by a 2nd drying step at 40 °C. The amount of lysozyme associated with the composites was quantified by determining the total carbon content in solids by mass spectrometry (DELTAplusXL ThermoFisher) with a Carlo-Erba NC2500. From these analyses the lysozyme content was calculated using the molecular formula C613H959N193O185S10 and the molecular weight of 14313 g/mol for lysozyme [66 (link)] (ProtParam based on UniProtKB entry P00698).
8
Stable Isotope Analysis of Gorgonian Symbiont
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For details of sample preparation please refer to the Supplementary Online Methods . In brief, each dried gorgonian fragment was physically homogenized, and the host and symbiont fractions were separated using centrifugation of a suspended homogenate. The supernatant (host) and pellet (symbiont) were dispensed into silver capsules, acidified and dried before stable isotope analysis.
All samples were analyzed using a Thermo Delta V isotope ratio mass spectrometer coupled to a Carlo-Erba NC2500 elemental analyzer via a Conflo III open-split interface at the Geophysical Laboratory. Analytical precision was determined by repeated analysis of an internal acetanilide standard (‘acet 6' 70% C). Standard precision during runs of initials and dark incubated enriched samples was +/− 0.2‰, and +/− 0.4‰ for the light incubated enriched samples.
All samples were analyzed using a Thermo Delta V isotope ratio mass spectrometer coupled to a Carlo-Erba NC2500 elemental analyzer via a Conflo III open-split interface at the Geophysical Laboratory. Analytical precision was determined by repeated analysis of an internal acetanilide standard (‘acet 6' 70% C). Standard precision during runs of initials and dark incubated enriched samples was +/− 0.2‰, and +/− 0.4‰ for the light incubated enriched samples.
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