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Sampling and analytical procedures:<\/strong><\/p>\n \u20281. Analysis of NO3- N and O isotope ratios with the denitrifier method<\/p>\n The denitrifying bacteria strains Pseudomonas chlororaphis <\/em>f. sp. aureofaciens<\/em> (ATCC 13985, Manassas, VA, USA) and Pseudomonas. chlororaphis<\/em> (ATCC 43928, Manassas, VA, USA) were used. Cultures were inoculated from cryo-preserved aliquots (Weigand et al., 2011) into sterile growth media prepared as originally described (Sigman et al., 2001; Casciotti et al., 2002) in 700 mL glass bottles containing 600 mL of medium, then sealed with gas-tight lids. Cells were cultured for 7-10 days at 20\u02daC on a rotary shaker table. Cultures were harvested by centrifugation and resuspended into 220 mL of fresh medium without potassium nitrate addition, achieving ca.<\/em> 3-fold concentration of the bacteria. Two mL of the cell concentrates were added to respective 20-mL headspace glass vials, capped with pre-rinsed butyl rubber septa and crimp-seals (Mcllvin and Casciotti, 2011). Vials were sparged with a water-scrubbed N2 gas stream for 6 hours to remove any N2O produced from the residual NO3- in the medium. NO3- samples were then injected into each vial to achieve a final sample size of 10 nmoles of N. Vials were incubated inverted in order to prevent potential N2O leakage. Following overnight incubation in the dark, ca.<\/em> 0.1 ml of 10 mol L-1 NaOH was injected into each vial to kill the cultures and sequester CO2 into carbonate species. The N2O gas in the vials was extracted, purified and analyzed with a Delta V Advantage continuous flow gas chromatograph isotope ratio mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) interfaced with a modified Thermo Fisher Scientific Gas Bench sample preparation device fronted by dual cold traps (Casciotti et al., 2002) and a GC Pal autosampler (CTC Analytics, Zwingen, Switzerland). Samples were referenced to pure N2O injections from a common reference gas cylinder.<\/p>\n 2. Demonstration of volume effects in analyzes of NO3- reference materials<\/p>\n The reference solutions were prepared from salts into primary stocks at 200 \u00b5mol L-1 in deionized water (DIW) from a Milli\u2010QTM water purification system (EMD Millipore, Burlington, MA, USA), and stored frozen. Primary stocks of NO3- reference materials (IAEA-NO3 and USGS-34) were diluted in NO3--deplete surface Sargasso seawater or in aged DIW to concentrations of 1, 5 and 20 \u00b5mol L-1, corresponding to respective injection volumes of 10, 2 and 0.5 mL, in order to aliquot 10 nmoles of N analyte. The NO3- aliquots were injected into the sparged bacterial concentrates of either P. chlororaphis<\/em> or P. aureofaciens<\/em>. Following bacterial conversion, the resulting N2O in the reaction vials was extracted, purified and analyzed on the isotope ratio mass spectrometer.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"Volume-dependent offsets in NO3- N and O isotope ratios of reference materials","@language":"en-US"}],"http:\/\/www.w3.org\/2000\/01\/rdf-schema#label":[{"@value":"Incidence of volume effect","@type":"xsd:string"}],"http:\/\/ocean-data.org\/schema\/hasProcessingDescription":[{"@value":" Processing notes from submitter:<\/strong><\/p>\n BCO-DMO processing notes<\/strong><\/p>\n