Quanta vs Invitrogen
qScript™ microRNA Quantification System Comparison to Invitrogen (hsa-miR-27a)
microRNA cDNA synthesis was performed starting with log-fold dilutions of human brain total RNA (from 1.0 g to 100 pg) using the qScript™ microRNA cDNA Synthesis Kit (Quanta) or the NCode™ VILO™ miRNA cDNA Synthesis Kit (Invitrogen). 1/100th of each microRNA cDNA reaction was used in a 20 µL qPCR amplification reaction specific for hsa-miR-27a using each manufacturers validated microRNA assay, qPCR mix and recommended protocol. Averaged plots for triplicate qPCR reactions for each input quantity are shown. Cycling conditions for the microRNA qRTPCR primer with NCode EXPRESS SYBR GreenER? qPCR SuperMix (Invitrogen) and PerfeCTa® MicroRNA Assay with PerfeCTa SYBR® Green SuperMix™ (Quanta) consisted of 95°C, 2 min followed by 40 cycles of 95°C, 15s; 60°C, 1 min.