Because these samples had one target gene CT value >30 and a CT value for the housekeeping gene
<30, [DELTA]CT values were positive and were located under these cutoffs in the samples with failed expression.
We report here an RT-PCR-based assay combining the determination of expression of both human IMPDH isoenzymes based on normalization to a housekeeping gene
Usually, the well-known housekeeping genes
are chosen such as the glyceraldehydes-3-phosphate dehydrogenase (GAPDH), [alpha]-actins (ACTB), hypoxantine phosphoribosyltransferase (HRPT), [beta]-tubulin (TUB), elongation factor 1 alpha (EF1A) and 18S, 28S rRNA (Shu et al.
The relative p53-like gene expression was evaluated using 18S rRNA as a housekeeping gene
Many housekeeping genes
do show different expression patterns under different experimental conditions and the influence of physical exercise on housekeeping gene
expression is not well studied (Jemiolo and Trappe, 2004; Mahoney et al.
The standard curves were prepared for both tested and housekeeping gene
, GAPDH (GenBank: U48832).
RNA integrity of the cDNA preparations was tested by real-time PCR amplification of the porphobilinogen deaminase housekeeping gene
LUC] yields and housekeeping gene
ratios), we used the nonparametric paired Wilcoxon test.
For each sample, the quality of DNA extraction was verified by real-time PCR for a housekeeping gene
encoding p-actin (7).
We also quantified the transcripts of the housekeeping gene
glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous control to normalize each sample using sense and antisense primers, 5' CCA TCA CCA TCT TCC AGG AGC 3' (positions 278-298) and 5' GGA TGA TGT TCT GGA GAG CC 3' (positions 663-682), respectively (Tokunaga et al.
ACTIN, a housekeeping gene
with expression unchanged during THH treatment, was chosen to serve as a control.
mRNA from both the gene of interest and a housekeeping gene