Figure 1. Rapid and sensitive amplification performance
Isothermal amplification of beta actin from human lung total RNA using IsoFast™ Bst 1-Step Mix. A primer mix of 0.2µM for F3 and B3 primers, 1.6µM for FIP and BIP primers, and 0.8µM for LoopF and LoopB primers were used. The total reaction volume was 25µL. 7 serial dilutions of template were used, corresponding to 250ng, 25ng, 2.5ng, 250pg, 25pg, 2.5pg, and 250fg of total RNA. The reaction was run at 65˚C for 34 minutes. A BioRad CFX96 Touch instrument was used to record fluorescence every 10 seconds. Time to result is the time required to reach the same fluorescent threshold. IsoFast™ Bst 1-Step Mix provides rapid and sensitive amplification down to 2.5pg of total RNA.