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Water was sampled from the CTD under trace metal clean conditions into 2 L polycarbonate bottles (acid-washed) and amended with 13C-bicarbonate. Flow cytometry samples were collected from triplicate bottles at several time points between 0 hours (unlabeled control) and 36 hours after the initiation of incubation. Flow cytometry samples were collected in 1 mL volumes and fixed immediately with 25% TEM-grade glutaraldehyde to a final concentrations of 0.125%. Samples were inverted 10 times to mix, incubated at room temperature in the dark for 10 minutes, then flash frozen in liquid nitrogen to archive. Samples were stored at -80C until analysis. For analysis, each sample was thawed at room temperature then analyzed by flow cytometry.\u00a0
\nCell counts were determined by flow cytometry using a BD Biosciences Influx high speed cell sorter. A 488 nm laser was used in addition to chlorophyll and phycoerythrin filters, forward scatter relative to 1 um beads, and side scattered light.\u00a0
Counts of Prochlorococcus from on-deck incubations with 13C-bicarbonate as part of DNA-SIP experiments; conducted on Hawaii Ocean Time-series (HOT) cruises 283 and 288.
The program BD Software was used\u00a0to collect flow cytometry data and generate distinct files for analysis. The program Flow Jo (version 7.6.5) was used to process flow cytometry files.
\nBCO-DMO Data Processing:
\n- modified parameter names to conform with BCO-DMO naming conventions;
\n- replaced spaces with underscores and removed parentheses\u00a0(in depth column).