The number of OTUs recorded per sample varied from only 103 for a gut sample of the nudibranch Phyllidia picta to 3704 for a sediment sample (Supplementary Data 1).
NTA post-acquisition settings were optimized and kept constant between samples, three videos 60 s long were recorded per sample and were analyzed to give the mean, mode, and median particles size together with an estimate of the number of particles.
Briefly, the E for each primer set was recorded per sample and an average E (EA) was then calculated for each primer set.
Labeled cells were sorted by fluorescence using a BD FACSCalibur Flow Cytometer (BD Biosciences) and a minimum of 30,000 events were recorded per sample using BD CellQuest Pro software.
At least 10,000 events were recorded per sample.
A total of 10,000 cell events were recorded per sample and data were analysed using FlowJo software (Tree Star, Ashland, OR).
Labeled cells were sorted by fluorescence using a BD Flow Cytometer (BD Biosciences) and a minimum of 10 000 events were recorded per sample using BD FACSDiva Software BD Biosciencess).
Subsequently, 10,000 events were recorded per sample with a FACScan flow cytometer (Becton Dickinson, Franklin Lakes, NJ) and analyzed using CellQuest software (Becton Dickinson, Franklin Lakes, NJ).
For H2AX phosphorylation quantitative analysis, foci were counted by eye until at least 40 cells and 40 foci were recorded per sample.
To ensure that all samples were within the optimal range of probe density for image analysis, we confirmed that there was no systemic increase in negative control counts as a function of total number of counts recorded per sample.
