Quantitative Real-Time RT-PCR

Quantification of gene expression by real-time RT-PCR has become the gold standard to which all other quantification methods are compared. Two very simple precautions, normalizing the cDNA to contain equal amounts of RNA and validation of the internal control gene, combined with RNA of sufficient quality, will almost certainly guarantee high quality data time after time. Described here is a protocol to normalize the RNA content of multiple cDNA reactions using the NanoDrop ® ND-1000 Spectrophotometer.