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The tube was thoroughly mixed by vortexing for 2 min.
Five (5) g of REG-80, REG-GB, and REG-FFA were suspended in 5 mL water, mixed with 5 mL chloroform, and then mixed by vortexing.
Briefly, 25 μL of an aqueous solution of H2O210%% (v:v) was added to 250 μL of 1 M NaOH in a microcentrifuge tube and mixed by vortexing.
A variety of solutions were then added to the tubes and thoroughly mixed by vortexing.
The supernatant was transferred to 60 µl of Tris-HCl buffer (pH 8.0) containing 40 µl of 10N NaOH and mixed by vortexing.
A phenolic detergent (500 µl, TRI Reagent, Molecular Research Center) was added, and the sample was mixed by vortexing for 20 sec and then incubated for 5 min at 25°C.
The lipid film was rehydrated in Tris-buffered saline (20 mM Tris, 150 mM NaCl, pH 7.4) containing 50 mM octyl-β-glucoside (Pierce) and thoroughly mixed by vortexing.
Then 200 μL CT-FCS was added and again the content was mixed by vortexing.
To this mixture, 1.5 μL Attractene was added, and each sample was mixed by vortexing.
Samples were dissolved in 200 μL of water and mixed by vortexing for 10 s.
Four hundred microliters of AL buffer (QIAGEN) was added to the cell lysate and mixed by vortexing for 10 s.
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