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Gently vortex the plate to mix.
Gently vortex the cells and incubate for 15 min at RT in the dark.
First, the discs for each study group were placed in a sterile plastic tube (Sarstedt, Norway) containing 25 mL saline and gently vortex mixed (MS2 Minishaker; IKA Works Inc., Wilmington, NC) at 100 rpm for 10 seconds.
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The tube was gently vortexed and incubated for 5 min at 4 °C in the dark.
All samples were gently vortexed and prepared in triplicate.
The solution was gently vortexed for 30 s and further incubated at 25 °C for 30 min.
The separated ECCN was also similarly suspended back in the aqueous buffer, gently vortexed and the superparamagnetic nanoparticles were separated by a magnetic separator.
Pre-cold lysis buffer (10 mM Tris-HCL, pH 7.4, and 0.03 mM PMSF) was added into the packed erythrocytes (volume ratio was about 40 1) and gently vortexed.
Afterward, bacterial cultures were centrifuged at 4000 rpm for 20 min, and 0.5 mL of the supernatant was added to 1 mL of Salkowski reagent (98 mL 35%% HClO4, 2 mL 0.5 M FeCl3) and gently vortexed.
The mixtures were gently vortexed and carefully transferred to a VC-H075P plain-type hematocrit capillary tube (Terumo Corp. Tokyo, Japan) with an RUVF-203S glass fiber UV irradiator (Radical Research, Tokyo, Japan, 200 W Hg-Xe lamp).
The plates were gently vortexed and resuspended in 200 μL of ammonium chloride buffer to lyse red blood cells, and the plates were incubated for 5 10 min at room temperature.
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