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To confirm that the correct amplicon size was obtained, standards and reactions were analyzed by agarose gel electrophoresis.
Peaks were identified using NIST08/Wiley09 combined spectra library, and some peaks were confirmed by retention times and spectra with commercially obtained standards (Lactic acid, Glycerol, Squalene).
To test the effect of the two candidate compounds identified in this work, commercially obtained standards were injected alone or in combination using the same method.
All observed pigments were also compared to commercially obtained standards of the free acids of biliverdin and protoporphyrin from Frontier Scientific Inc. (UT, USA) dissolved in acetonitrile-acetic acid.
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By using the reference molecule, we obtained standard curves with good linearity and relatively high efficiency of PCR reaction.
All experiments were performed twice and the obtained standard deviation was always less than 5%% of the reported value.
Concentrations of naringenin were then extrapolated from the previously obtained standard curve (Figure 1C).
The concentrations were calculated by the optical density measurements from the obtained standard curve.
We obtained standard radiographs of the lumbar and thoracic spine at baseline and follow-up.
We obtained standard 2D parasternal long and short-axis to determine left ventricular dimensions.
We obtained standard reference material DEP from the National Institute of Standards and Technology (Gaithersburg, MD, USA).
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