FAQ: Is the Monarch® Total RNA Miniprep Kit compatible with Luna® RT-qPCR Reagents?

Yes. RNA purified with this kit can be used for RT-qPCR with NEB’s Luna One-Step RT-qPCR reagents.

RT-qPCR of Monarch-purified RNA purified from various sample types consistently produces high-quality results, allowing accurate and reproducible quantitation

dots in boxes
To demonstrate the quality and utility of Monarch Total RNA for RT-qPCR-based quantitation, 16 experiments were performed and visualized using the “dots in boxes” approach developed at NEB. Each experiment encompasses triplicate reactions for a five-log range of input template concentrations and no-template controls (NTC) (18 reactions total). A high-quality RT-qPCR experiment, represented by a dot inside the box, is defined as having an efficiency between 90-110%, a ΔCq of ≥3 (where ΔCq is the difference between the average Cq of the NTC and the lowest input), and a Quality Score ≥4 (based on linearity, reproducibility, curve shape, and other parameters). Primers targeting GAPDH variants (rat, mouse, tomato, yeast), PGK variants (rat), and B-actin variants (rat, mouse, tomato) were used with total RNA purified from rat brain, rat liver, rat kidney, mouse kidney, tomato leaf, and yeast as templates. RT-qPCR assays were assembled as directed using Luna RT-qPCR reagents, and cycled on a Bio-Rad CFX384.

 

RT-qPCR on Monarch-purified RNA generates high-quality qPCR curves demonstrating accurate quantitation across a wide variety of sample types

RNA qPCR
Monarch-purified RNA from human whole blood, rat kidney, tomato leaf, and the yeast S.cerevisiae was diluted to produce a five-log range of input template concentrations. Primers targeting GAPDH variants (tomato, yeast), PGK (rat), or SMG1 (human blood) were used for RT-qPCR assays, assembled as directed using Luna RT-qPCR reagents (NEB #E3005) and cycled on a Bio-Rad CFX384.