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The supernatant was carefully removed, and concentrations of each sample was determined using a bicinchoninic acid (BCA) assay.
After adding 4 mL of millipore-filtered deionized water and briefly centrifuging, the mineral element concentrations of each sample were determined by atomic absorption spectrophotometer (SHIMADZU AA-6300) (Zhang et al. 2011).
For these statistical analyses the data (amino acid concentrations of each sample) were log-transformed.
Protein concentrations of each sample were determined using the method described by Lowry et al [49].
After clearing the lysates by high-speed centrifugation, protein concentrations of each sample were determined using a Protein Bradford assay Bio-Rad Laboratories AGG, Reinach, Switzerland).
1ug of DNA-free RNA was run out on a non-denaturing gel to ensure equal concentrations of each sample followed by reverse transcription of 1 µg of each RNA sample using the iScript cDNA synthesis kit, following manufacturer's instructions (BioRad, Hercules, CA).
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The protein concentration of each sample was determined with a Micro BCA Protein Assay Kit (ThermoScientific).
The concentration of each sample was determined with UV spectroscopy by using the standard curve method (λmax, 484 nm).
The DNA concentration of each sample was quantified using Qubit dsDNA BR Kit according to the manufacturer's instructions (Thermo Fisher Scientific).
Finally, absorbance values were calculated for standards and samples and the concentration of each sample was determined using the standard curve.
The concentration of each sample is 0.1 wt%.
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