Choice of primer location for qPCR from AA-aRNA. One μg of universal reference RNA and 100 ng AA-aRNA were reverse transcribed using our optimized RT protocol. Resulting cDNAs were diluted 10-fold and subjected to qPCR using 5 pairs of primers located at different distances of the 3' end of the hif1a gene. Cq values were plotted against these distances. Linear regression lines illustrate the effect of increasing the distance from 3' end. Moving away primers from the 3' end increased Cq values when performing qPCR with cDNA generated from AA-aRNA.
Jeanty et al. BMC Genomics 2010 11:542 doi:10.1186/1471-2164-11-542