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We used two analysis methods, geNorm and Norm-Finder, to identify the best combination of reference genes and then correlated housekeeping gene expression with DNA content and gel representation of ribosomal RNA.
Additional File 3 shows the results obtained from the Bestkeeper© analysis to select the best combination of reference genes used for the normalization of the qRT-PCR data.
With the use of the best combination of reference genes (miR-16 and miR-345), significant dysregulation of all four target miRNAs (miR-21, miR-31, miR-143 and miR-145) was detected.
NormFinder also estimates the best combination of reference genes by sub-grouping in 'all stomach tissues' - RPL29-B2M (0.005) and in 'all stomach cell lines and tissues' - 18S rRNA-B2M (0.013).
The values of Cq presented here should not be considered alone, but they may help in the selection of best combination of reference genes when there is previous data about target gene expression levels.
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