Figure 5

Side-by-side quantification of let-7b in human total RNA samples isolated from the cell lines: A549, HeLa, and HT-29 in two independent qPCR runs. Experiments were performed using the miR-Q approach as well as the mirVana™ qRT-PCR Detection Kit. For this purpose, total RNA samples (50 ng/μl) were either spiked with 100 pM synthetic let-7b (A549 si, HeLa si, and HT-29 si) or remained non-spiked (A549, HeLa, and HT-29). RT reactions were performed with 50 ng, 25 ng, and 5 ng of all RNA samples, followed by qPCR detection of let-7b in different runs. Columns represent the mean (± SD) of three measurements. A) let-7b quantification by means of the miR-Q approach using both the spike-in controls and the non-spiked samples. B) Quantification of let-7b by means of mirVana™ qRT-PCR using both the spike-in controls and the non-spiked samples.