Figure 3. Rapid and consistent amplification performance across a range of templates
Isothermal amplification of 3 targets using IsoFast™ Bst Mix, NEB WarmStart® LAMP Kit, and NEB Bst 2.0 DNA Polymerase. The manufacturers’ protocols were followed to set up the reaction mix. The primer mix consists of 0.2µM for F3 and B3 primers, 1.6µM for FIP and BIP primers, and 0.8µM for LoopF and LoopB primers. The total reaction volume is 25µL. 7 serial dilutions of template were used (8 for scaffolding protein gene), corresponding to the copy number indicated. The reaction was run at 65°C for 100 minutes. A BioRad CFX96 Touch instrument was used to record fluorescence every 10 seconds. The time to result is the time required to reach the same fluorescent threshold. IsoFast™ Bst Mix shows fast amplification and consistent performance across a range of templates when compared to leading manufacturers.