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Expression of reference gene(s) is expected to remain unchanged in fluoride-treated cells or in rodents relative to the corresponding untreated controls.
NormFinder is another approach to assess the stability of expression of reference genes [ 14].
Abundances of the 23 unigenes were calculated relative to the expression of reference genes DlFSD1a, EF-1a, and eIF-4a.
Results were normalized against the expression of reference genes ODC and RPL8 using CFX Manager software (BioRad).
Both problem and calibrator samples were normalized by the relative expression of reference small non-coding RNAs (SNORD44 and SNORD48).
In the pig however, very little is known about the expression of reference genes, certainly in combination with real-time RT-PCR.
Similar(41)
We measured the expressions of reference (18S) and target genes in separate tubes.
The relative expression levels were calculated based on the expression level of reference genes by qBase (Hellemans et al. 2007) and data were analyzed with SPSS 19.0 software (IBM, Chicago, IL).
Thus, for each examined gene, the expression levels of the examined gene in the apical/basal cell samples were normalized to the expression levels of reference genes, GAPDH/Polyubiquitin/Ntubc2.
In the pig however, very little information is available on the expression stability of reference genes.
However, expression instability of reference genes in tumors may introduce bias when determining miRNA levels.
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