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Another change in the process required for scaling up is the replacement of cell lysis by sonication for high pressure disruption or by chemical methods.
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Lyse cells by sonication for 60 sec on a Sonic Dismembrator (Fisher Scientific) with macrotip probe.
All filters were extracted in 15 mL of high-purity methanol (LC-MS CHROMASOLV-grade, Sigma-Aldrich) by sonication for 45 min. The methanol extracts were then blown dry under a gentle N2 stream at ambient temperature.
For MeHg, homogenized fish sample was extracted with 50% HCl by sonication for 3 h.
The resulting FGO/PS solution was stirred for 2 h followed by sonication for 30 min.
All of the extractions were facilitated by sonication for 3 × 20 min.
Then, cells were disrupted by sonication for three 5 min intervals.
Cells were then lyzed by sonication for 30 minutes followed by centrifugation at 20 500 rpm for one hour.
Solubilization was attained by sonication for 3 × 40 s.
For this assay, hydrophobic coverslips were prepared according to the following scheme: sonication in acetone for 15 min was followed by sonication for 15 min in ethanol.
Cells were incubated on ice for 10 minutes, followed by sonication for 15 seconds.
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