Sediment and porewater handling:

PCG02-08- Core material was sectioned and pressure-squeezed with a modified Reeburgh squeezer (Reeburgh, 1967) into acid-washed all-plastic syringes. Porewater was then filtered through 0.45 um polyethersulphone syringe filters.

Hydrates2004- Core material was sectioned and pressure-squeezed with a modified Reeburgh squeezer (Reeburgh, 1967) into acid-washed all-plastic syringes. Porewater was then filtered through 0.2 um arodisc polyethersulphone syringe filters.

AT50-14- Core material was sectioned and pressure-squeezed with a modified Reeburgh squeezer (Reeburgh, 1967) into acid-washed all-plastic syringes. Porewater was then filtered through pre-combusted 0.4 um glass fiber filters.

AT50-29B- Core material was sectioned into acid-washed falcon tubes and refrigerated. Porewater was extracted using Rhizons with a 0.2 um filter size.

Porosity: Fresh sediment was placed in pre-weighed petri-dishes, sealed, and refrigerated until measurement in lab. Petri dishes were weighed upon return to shore, then were played in a drying oven (45℃, 1 week). Petri dishes with dried sediment were weighed again. The difference between the wet and dry sediment is the water weight, and the ratio of the water volume to the total volume is the porosity, assuming a dry bulk density of 2.5.

SO4_mM: 0.5 mL porewater was sub-sampled into 2mL microcentrifuge tubes and dosed with 50 uL 0.1M phosphoric acid. Samples were measured using ion chromatography. IAPSO certified seawater was used for standardization, and precision is ±1.5%.

CH4_uM: Sediment plugs (volume = 3-6 cm3) were sampled and stored in serum bottles sealed with 1-cm-thick butyl rubber septa. 5mL of saturated brine was added to the serum bottles and shaken to equilibrate the headspace. The samples were refrigerated until analysis. For cruises PCG02-08, Hydrates2004, and AT50-14, methane concentrations of serum bottle headspaces were determined by gas chromatography flame ionization detection. These headspace concentrations were used to calculate dissolved concentrations of methane in porewater. For samples measured by GC, dissolved concentrations were calculated following Hoehler et al. (2000).

For cruise AT50-29B, methane concentrations were determined using cavity ringdown spectroscopy. Check standards for concentration were run approximately every 5 samples and corrected as described by Pohlman and others (2021). For samples measured by cavity ring-down spectroscopy, dissolved concentrations were calculated using the following equation:

((pCH4 * 10^-6 * Vg) / (R * T_extraction) + (Vw * pCH4 * 10^-6 * Sol_CH4)) / Vw * 10^6 = [CH4]

Where pCH4 is the headspace concentration of methane in ppm, Vg is the volume of gas sample in mL, R is the gas constant in L-atm/mol-K, T_extraction is the extraction temperature in Kelvin, Vw is the volume of water sample that was extracted in mL, Sol_CH4 is the solubility of methane in mol/L-atm (Wiesenburg & Guinasso, 1979; Yamamoto et al., 1976), and [CH4] is the dissolved concentration of methane in the original natural water sample, in micromoles per liter.

DOC_uM and DOC_d13C_permille: Samples (2-4 mL) were stored in combusted borosilicate glass vials with acid-cleaned teflon-lined septa caps. Vials were pretreated with 20 percent hydrochloric acid (12 microliters per milliliter of sample) to decrease the pH of the sample to 2 or less, and samples were stored at 4 degrees Celsius until analysis. Concentrations were determined with a TOC Analyzer using wet chemical oxidation (cruises PGC02-08 and Hydrates2004) (Heuer et al., 2009; Pohlman et al., 2011) and high temperature catalytic oxidation (cruises AT50-14 and AT50-29B) to convert DOC to CO2. For all cruises, nondispersive infrared detection was used to quantify this CO2. Potassium hydrogen phthalate (KHP) calibration standards were run to quantify DOC concentration. Stable carbon isotopes (d13C-DOC) were measured by isotope ratio mass spectrometry. Stable carbon isotope ratios are reported in standard delta notation relative to Vienna Pee Dee Belemnite (VPDB). Quality assurance included replicate analysis of natural reference materials, field samples and calibration standards. Uncertainty in DOC concentration was 5 percent or less, and uncertainty in d13C was 0.2 permille or less, based on replicate measurements.

DIC_mM and DIC_d13C_permille: Samples for DIC concentration for cruises PCG02-08 and Hydrates2004 were collected as 2 mL aliquots in 5 ml serum vials, sealed with butyl rubber septa, and quantified by coulometry. Samples for stable carbon isotopes (d13C-DIC) for cruises PGC02-08 and Hydrates2004 were collected as 1 ml aliquots in 2 ml serum vials, sealed with butyl rubber septa and analyzed as CO2 by isotope ratio mass spectrometry following acidification of the sample (Pohlman et al., 2013). Samples for DIC concentration and stable carbon isotopes for DIC for cruises AT-50-14 and AT50-29B were collected by injecting 1mL porewater into a helium-filled 12 mL exetainer vial pre-filled with 1mL 85% phosphoric acid. The stable carbon isotope ratios and concentrations were determined from CO2 by isotope ratio mass spectrometry. Measurements are standardized with lithium carbonate reference material, and isotope ratios are reported in the standard d-notation relative to VPDB.

TOC_wt.% and TOC_d13C_permille: Sediment samples from all cruises were freeze-dried and ground with a mortar and pestle. Homogenized sediment was weighed into silver capsules for acid fumigation. Samples were acid fumigated to remove carbonate and dried at 45℃ for 24 hours, then wrapped into tin capsules. Samples were run on an elemental analyzer interfaced with an isotope ratio mass spectrometer. Samples from PGC02-08 were normalized to an acetanilide standard and samples from AT50-14 were run with a peptone reference material, as well as standards USGS 40 226 and USGS 41. Stable carbon isotope values are reported in d notation relative to VPDB.

H2S_mM: Porewater sulfide (ΣH2S) was determined by sparging the sample aliquot after acidification with 25 % phosphoric acid, and trapping evolved H2S in sulfide antioxidant buffer (SAOB) solution for measurement with a sulfide specific electrode. Standards were prepared from a sodium bisulfide stock solution that was titrated with lead nitrate to determine its concentration, daily. Also on a daily basis, the stock solution was serially diluted with SAOB solution to produce a five-level calibration.