Quantitative Real-Time PCR for Adenovirus-Infected Cells
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Corresponding Organization :
Other organizations : National Institutes of Health, National Heart Lung and Blood Institute, National Institute of Arthritis and Musculoskeletal and Skin Diseases
Variable analysis
- Adenovirus infection of cells
- Gene expression levels
- RNA isolation method (PureLink RNA Mini Kit)
- Reverse transcription reaction (oligo(dT)20 and SuperScript III First-Strand Synthesis System)
- Real-time PCR reaction conditions (10 μl total volume, 2 μl cDNA, 1 μl gene-specific primer mix, 5 μl SYBR GreenER qPCR SuperMix, 2 μl water)
- Real-time PCR thermal profile (50°C for 2 min, 95°C for 10 min, 35 cycles of 95°C for 15 s and 60°C for 1 min)
- Reference endogenous gene (GAPDH) for normalization
- RNA from cells infected with control adenovirus (Ad. Null)
- Not explicitly mentioned
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