Quantification of mRNA Levels by Real-Time PCR
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Corresponding Organization : University of Wisconsin–Madison
Protocol cited in 2 other protocols
Variable analysis
- Real-time PCR protocol
- Reverse transcription with Oligo (dT) primer
- Amplification with gene-specific primers and Power SYBR Green PCR Master Mix
- Use of Step-1 Real-Time PCR System
- MRNA levels of different samples
- Amount of cDNA used for each PCR reaction (1 µg, except 5% of cDNA for IP samples)
- Use of Universal Master Mix (Applied Biosystems)
- Final reaction volume of 20 µL
- Input samples for RIP-real time PCR (used to normalize IP samples)
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