IsoFast Hot Start Bst Polymerase Colour and Mix

Reliable colourimetric readout with cold and ambient reaction setup
Isothermal amplification of a target sequence in the scaffolding protein gene from the M13 bacteriophage genome using IsoFast™ Hot Start Bst Polymerase or IsoFast™ Bst Polymerase in 10x IsoFast™ Colour Buffer A, and NEB WarmStart Colorimetric LAMP 2X Master Mix. A primer mix consisting of 0.2 µM for F3 and B3 primers, 1.6 µM for FIP and BIP primers and 0.8 µM for LoopF and LoopB primers was used. The total reaction volume was 25 µL. 8 serial dilutions of M13 ssDNA genome were used, starting with a stock of 0.5 ng/µL and using a dilution factor of 10, corresponding to the number of genome copies indicated next to the plates. Reaction master mixes and plates were prepared either using cold blocks (cold Setup) or at room temperature (ambient setup), for approximately 20 min. The reaction was run at 65 °C for 30 minutes. Plates were then photographed to show the colours obtained at the end of the run. IsoFast™ Bst Polymerase and IsoFast™ Hot Start Bst Polymerase showed a better sensitivity compared to NEB WarmStart Colorimetric LAMP 2X Master Mix in Cold Setup.
IsoFast™ Hot Start Bst Polymerase allows easy screening for positives even with ambient temperature setup.