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Additional file 3: Evaluation of protein quantification; The standard sample was combined at a ratio of 0.5 1 2 by volume.

Immediately after HPLC analysis, 0.80 mL of the filtered sample was combined with 0.25 mL of 5 g/L Ferrozine (Acros, 99.9%) and 3.25 mL 40 mM, pH 3.75 acetate buffer.

For P2, the isolated DNA from the squeezed sample was combined with total DNA isolated from the whole L. patella sample (1∶1 mass ratio).

One µg of each RNA sample was combined with loading buffer, and gels subject to electrophoresis at 80 V for 40 50 min at room temperature.

Each tissue sample was combined with 1 ml Trizol (Invitrogen, Carlsbad, CA) and a 5 mm steel bead and homogenized on a TissueLyser (Qiagen, Valencia. CA) for 4 min at 30 Hz, rotating the assembly halfway through the time.

Each sample was combined with 500 µl of 2x buffer B (143 mM Tris pH 8.0, 143 mM NaCl, 14 mM EDTA and 5.7% SDS), 500 µl phenol chloroform, and approximately 0.25 g zirconium beads (0.1 mm, Biospec Products Inc).

Reflected light from the tissue sample is combined with reflected light from the reference.

Whenever different channels of the same sample were combined for analysis, the read counts were summed prior to normalization.

PCR products for each sample were combined and purified using a PCR cleanup kit (Invitrogen corp., Carlsbad, CA).

All three extracts for each sample were combined and lyophilized.

RNA extracts from the same sample were combined.