RT protocol optimization decreases qPCR Cq values. Total RNA extracted from blood cells of the same 15 brain-dead organ donors used in Figure 1 was amplified and amino allyl labeled to generate AA-aRNA. 100 ng of each of these AA-aRNA were then reverse transcribed to cDNA using either the original or the optimized RT protocol, and qPCR was applied to assess expression levels of ACTB, GAPDH, LY96, ELANE, ANXA1, STAT3 and TNF-α. Results are represented as Cq values for each gene (mean ± SD of the 15 donors) and as the mean ± SD of the 7 genes. The optimized RT protocol decreased Cq values. ND: not detectable. * P < 0.001 and # P = 0.004 vs original RT protocol (paired t-test).
Jeanty et al. BMC Genomics 2010 11:542 doi:10.1186/1471-2164-11-542