Description
Loop-mediated Isothermal Amplification is a powerful DNA amplification method that works at a constant temperature (usually 60ºC-65ºC) without the need of a thermal cycler. It achieves extremely rapid and specific amplification (typically 10⁹–10¹⁰ copies in 15–60 minutes) using 4 to 6 specially designed primers and a strand-displacing DNA polymerase.
proACE™ Bst Visual Mix is a 2x Mastermix for colorimetric Loop-mediated Isothermal Amplification. It comprises proACE™ Hotstart Bst Polymerase (the large fragment of Bacillus stearothermophilus DNA polymerase, expressed in E. coli, combined with a proprietary hot start technology). This large fragment of Bst DNA Polymerase preserves 5´→3´ polymerase activity for DNA synthesis but is devoid of 5´→3´ exonuclease activity, thereby facilitating the strand displacement mechanism that is critical to Loop-mediated Isothermal Amplification. The reversible hot start technology allows for setting up the experiment at ambient temperature, reducing non-specific amplification that may occur below 45ºC, which is ideal for point-of-care and field testing. In this particular mastermix, proACE™ Hotstart Bst Polymerase is supplied in a pH-based colored solution, which switches from orange (negative) to yellow (positive) in case of target amplification.