Table 1 Determination of Coefficients of Variation (CV) to examine variability in transcript numbers for different NT/CT ratios: The CV between the initial ACTB transcript number in the NT was calculated based on the observed NT/CT ratios at the end of StaRT PCR amplification. LD419 cDNA was mixed with CT to obtain assigned NT/CT ratios: 0.1, 0.5, 1, 5, and 10. Independently, each of these 5 assigned NT/CT ratios was used to set up separate reactions, which were subjected to PCR amplification. All experiments were performed in triplicate. The initial number of ACTB transcripts at each assigned NT/CT ratio was calculated at the end of the StaRT PCR amplification. CV between the calculated ACTB transcript numbers at the different NT/CT ratios was found to be the least when the ratio was kept close to 1. Further, the CV between the calculated ACTB transcript numbers was fairly low over a wide range of NT/CT ratios.

From: Sensitivity and reproducibility of standardized-competitive RT-PCR for transcript quantification and its comparison with real time RT-PCR