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The reproducibility of the assay was determined based on the Ct values of the amplified products as well as in BK copies per milliliter of sample.
One milliliter of sample was mixed with 4 mL of ethanol and DPPH solution.
One milliliter of sample solution was put into a test tube, then 1 mL phenol solution (5%) and 5 mL sulfuric acid were added and test tubes were shaken for 30 s.
One milliliter of sample aliquot from this bio-transformed product was drawn after 24, 48, and 72 h postincubation with silver nitrate solution, and absorbance was recorded by using Hitachi U-2000 spectropHitachier (Hitachi, Ltd., Chiyoda-ku, Japan) range between 350 and 600 nm in order to study the change in light absorption of the solution with increase in color intensity.
To analyze DNA content, cells were stained with a 0.0001% propidium iodide (Sigma) solution (1000X stock generated by suspending 13 mg PI into 8.6 ml H2O. Prior to use, this stock is diluted 10 ul +990 ul of Tris solution for each milliliter of sample).
The results were expressed as lg IFU per milliliter of sample.
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One milliliter of samples were taken after 30 min from the experimental tubes, added to ice cold PBS and centrifuged.
Four milliliters of sample solution was added to 1 ml reagent A. To this mixture 1 ml of reagent B was added.
Five milliliters of sample were collected periodically over an 8 hour time period, and then filtered through a 0.45 μm disposable filter into a 20 ml polyethylene scintillation vial containing 5 ml 10% HNO3 for sulfur and iron analysis.
Notably, plasmonic MS can be advantageous in terms of speed (reaction free, in minute overall analysis) and sensitivity (~ 6 nL of sample required), compared to the biochemical method that requires several hours for analysis (including bio-reactions and pre-treatment steps) and utilizes milliliters of sample.
Forty milliliters of sample was loaded each time.
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