Detailed methods used for qRT-PCR have been described previously (28 (link)). Specific primers were purchased from Qiagen (QuantiTech Primer Assays), for each primer set the efficiency was > 95% and a single product was seen on melt curve analysis. Relative expression levels were calculated using the 2-ΔΔCt method normalizing to GAPDH expression levels for each treatment and the fold increase in expression was relative to the smallest expression level.
Protocol detail
Efficient qRT-PCR Quantification Protocol
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Assays
Gapdh
Primer
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Corresponding Organization : Instituto Murciano de Investigación Biosanitaria
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Variable analysis
independent variables
- Specific primers were purchased from Qiagen (QuantiTech Primer Assays)
- Relative expression levels
- GAPDH expression levels
- The efficiency of each primer set was > 95% and a single product was seen on melt curve analysis.
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