Ten-fold serial dilutions of the RNA external calibrator were done in CHIKV-negative plasma, resulting in concentrations ranging from [10.
3 33 (a) Ten-fold serial dilutions of the RNA external calibrator were done in CHIKV CHIKV-negative plasma (copies/mL), submitted to RNA extraction, and tested in triplicates in the same routine run to evaluate the intraexperimental reproducibility.
After each block of six sample specimens, one external calibrator was measured.
Thus, the ratios of tryptophan per nitrotyrosine and kynurenine per nitrotyrosine for the external calibrator were calculated as well as for the sample, leading to the final results.
The company Roche (30) advises their customers working with relative quantification and external calibrators
to use a common calibration curve for any target as long as the amplification efficiency does not differ more than 0.
External calibration is based on a set of external calibrators containing elements of interest and internal calibrators in a simple acid.
The most widely used calibration method is by a set of external calibrators containing elements of interest and internal calibrators in a simple acid.