Validation of microarray results by semi-quantitative RT-PCR revealed temporal variation in gene expression profiles.
Fig. 4 Validation of microarray results by semi-quantitative RT-PCR.
Although moderate-to-good agreement has been reported between the two methods, validation of DNA microarray results by the more sensitive PCR array is generally recommended [38].
First, we explored the microarray results by comparing gene expression profiles among Cr_large, Cr_small and untreated control groups.
Although not shown in figure S1, another gene confirming microarray results by QRT-PCR was the antimicrobial peptide gambicin.
To validate the microarray results by an independent method we carried out qPCR experiments on homogeneous pools of 50 fibers.
We validated the microarray results by RT-PCR analysis in NUGC3 cells after SOX2 over-expression, and representative results are shown in Figure 6A.
These different controls exclude PCR reaction problems as an explanation for the lack of reproducibility of our microarray results by qPCR.
We confirmed our microarray results by quantitatively analyzing the expression of numerous chemokine ligands and receptors, using RT-PCR arrays (Fig. 5).
Validation of the microarray results by qRT-PCR showed a high Pearson's correlation coefficient (0.998) between the microarray and qRT-PCR data (Fig. 6).
Due to the low amounts of total RNA obtained from primary neuronal cells we chose to validate our microarray results by quantitative PCR using individual TaqMan assays on independent preparations of Aβ42-treated primary cultures.
