Epichlorohydrin-crosslinked α–, β–, and γ–Cyclodextrin prepolymer (CycloLab R&D) were used as received. The following items were purchased from Fisher (Waltham, MA, USA) at reagent grade and used as received: acetone; 1,6 diisocyanatohexane (HDI); dimethylformamide (DMF); hexanes; light paraffin oil; methylsulfoxide (DMSO); potassium hydroxide (KOH). MNC was purchased from Research Products International (Mt. Prospect, IL, USA).
Light paraffin oil
Manufactured by Thermo Fisher Scientific
Sourced in United States
Light paraffin oil is a transparent, colorless liquid used as a lubricant and insulating medium in various laboratory equipment and instruments. It has a low viscosity and is commonly used to prevent evaporation and maintain sample integrity in laboratory settings.
Automatically generated - may contain errors
4 protocols using light paraffin oil
1
Crosslinked Cyclodextrin Polymer Synthesis
2
Synthesis of Cyclodextrin Microspheres
3
Quantifying Syringafactin Production in Pseudomonas
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Atomized oil assay was used to detect syringafactin as previously described (Burch et al., 2010 (link)). PstDC3000 and all rsmA mutants were grown on KB agar plates for 48 h, resuspended in PBS, and adjusted to an OD600 = 1.0. Ten microliters was pipetted onto the surface of KB plates (1.5% agar) and incubated for 24 h at 20°C. An airbrush (VIVO HOME, Pleasanton, CA, United States) was used to spray a mist of mineral oil over the plates (light paraffin oil; Thermo Fisher Scientific, Waltham, MA, United States). Brighter oil drops formed a visible halo around bacterial colonies. The ring area of the halos was measured to represent syringafactin production. Experiments were performed in triplicate and repeated three times. Statistical comparison among different strains was performed using one-way ANOVA followed by Fisher’s LSD test (p < 0.05).
4
Biosurfactant Detection and Cell Hydrophobicity
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Biosurfactant detection assay was performed using an atomized oil assay as previously described, with a few modifications (Burch et al., 2010 (link)). Bacteria grown overnight were spotted onto LB or MM agar plates and incubated at 28°C for 48 h. A fine stream of light paraffin oil (ThermoFisher Scientific, Waltham, MA, USA) was applied onto the plate using an airbrush with an air pressure of 1055 g/cm2 (master airbrush model G22; TCP Global Co., San Diego, CA, USA). Halos of the biosurfactant were visualized and imaged immediately after spray. Oil droplets were observed using the Leica Zoom 2000 stereomicroscope (Leica microsystems, Wetzlar, Germany).
Cell surface hydrophobicity was measured using the hexadecane partitioning method (van Loosdrecht et al., 1987 (link)). Bacterial cells from overnight LB cultures were harvested and washed three times with 0.5× PBS buffer. Cells were then resuspended in 1 ml 0.5× PBS buffer and the OD540 values were determined. The 250 μl of n‐hexadecane (Sigma‐Aldrich, St. Louis, MO, USA) was then added to each cell suspension, and the suspensions were vortexed for 10 min. The resulting mixtures were incubated at 37°C for 30 min. The OD540 of the aqueous layer was measured against a blank of hexadecane‐extracted PBS. Cell surface hydrophobicity was calculated as follows: cell surface hydrophobicity (in %) = 100× (final OD/initial OD).
Cell surface hydrophobicity was measured using the hexadecane partitioning method (van Loosdrecht et al., 1987 (link)). Bacterial cells from overnight LB cultures were harvested and washed three times with 0.5× PBS buffer. Cells were then resuspended in 1 ml 0.5× PBS buffer and the OD540 values were determined. The 250 μl of n‐hexadecane (Sigma‐Aldrich, St. Louis, MO, USA) was then added to each cell suspension, and the suspensions were vortexed for 10 min. The resulting mixtures were incubated at 37°C for 30 min. The OD540 of the aqueous layer was measured against a blank of hexadecane‐extracted PBS. Cell surface hydrophobicity was calculated as follows: cell surface hydrophobicity (in %) = 100× (final OD/initial OD).
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