This was done by preparing serial dilutions from 10−1 to 10−6 by transferring 1 ml homogenized sample (1 10 dilution) to 9 ml sterile saline water, mixing well with vigorous shaking and continuing until 10−6 dilution was reached.
The dip-coating of the catalyst on the SiC foam was done by preparing slurry of definite quantity of catalyst powder and dipping the foam into that for a particular time period in the dip-coater.
Intra-day and inter-day precisions were done by preparing and applying three different concentrations of standard in triplicate six times a day and similarly on six different days, respectively.
The tensile testing of the material was done by preparing test specimens as per ASTM E8 (American Society for Testing and Materials, West Conshohocken, PA, USA) specifications, as shown in Figure 4a.
This was done by preparing a conventional wet mount (~12 μl of culture) and placing a drop of acid (~25 μl) at the edge of the cover slip to be drawn under by capillary action to react with potential carbonate minerals within the biofilms [see Additional file 1].
Activation of the proPO system in crayfish after bacterial challenge was done by preparing hemocyte lysate (HLS) and then determined LPS-activated PO activity in the HLS samples.
Interday and intraday precisions were done by preparing and applying three different concentrations of standards (in triplicate) on the same day and on three different days, respectively.
This was done by preparing an FITC-labeled G3-C12 peptide (G3-C12-FITC) and showing that it selectively accumulated in the kidneys soon after injection, probably due to reabsorption of the peptide by proximal renal tubular cells.
This was done by preparing a solution of concentrated red dye (0.05 mL of DMSO with 0.0015 g of Tetramethylrhodamin-5-isothiocyanate; T0820-5MG by Sigma-Aldrich), which was homogenized in PBS to make the diluted dye (1 μL of concentrated solution with 10 mL of phosphate buffered saline).
The labelling was done by preparing 40 mM G2M5 in 97 % H2O (Sigma Aldrich, Schnelldorf, Germany) and incubating at 90 °C for 30 min, which resulted in ~90 % O-labelling as deduced from the areas and intensities of the O- and O-labelled peaks in the MALDI-TOF MS analysis.
