Additionally, we checked the relative quantification of genes encoding IAA9 and PR1 in Citrus plants, by RT-qPCR, using RNA extracted from a mixture of leaves and petioles of Ponkan mandarin and Pera sweet orange at 1, 7, 14, and 21 days after inoculation of X. fastidiosa or mock inoculated (control).
We carefully selected five reference genes for accurate quantification of gene expression by RT-qPCR and checked the stability of their expression.
For each pair, four observed ratios of long to short products, including two for baseline correction and two of crude ratio between cell lines, were used to cross-check the accuracy of final quantification.
Preliminary validation of the method was performed by checking the linearity, precision, recovery, detection and quantification limits, and repeatability.
The possible osmotic role of glucose and urea is easily checked by their quantification in the urine.
After checking the UV spectra and determining workable wavelength, the samples were subjected to HPLC experiments using porous silica and core-shell columns for separation, identification, and quantification.
Bias Correction and Background Correction was checked for protein quantification and normalization.
Starting on Oct. 7, the second part of Phase Two--the quantification segment--begins.
Data was analyzed using "Label-free quantification" checked, and the "Match between runs" interval was set to 2 min.
All stainings were checked by a pathologist, who confirmed the quantification results as generated by the researcher as well.
