Fluorescent Dye

IsoFast Bst Polymerase is a recombinant protein expressed in E. coli and represents the large fragment of Geobacillus stearothermophilus (formerly known as Bacillus stearothermophilus) DNA Polymerase. This portion of the protein catalyzes the 5?-3? synthesis of DNA and has strand displacement activity but does not contain the 5?-3? exonuclease domain.1. Strand displacement refers to the ability of an enzyme to dissociate the hydrogen bond of double stranded template DNA encountered downstream, essentially unzipping the DNA as the complementary strand is synthesized. IsoFast Bst Polymerase displays strong strand displacement activity and is suitable for amplification methods including whole genome amplification, multiple displacement amplification and isothermal amplification. DNA synthesis is performed at a constant temperature, and we recommend running the reaction at 65 ?C. However IsoFast Bst Polymerase works well over a broad temperature range (55 ?C to 70 ?C, see Figure 4), giving a wider range of reaction conditions in order to optimize primer annealing and strand displacement. The enzyme is heat inactivated at 80 ?C. Designed for fast amplification speed, IsoFast Bst Polymerase gives rapid and consistent results across different target sequences (see Figure 1, 2 & 3). The enzyme is provided with an advanced 2-part buffer system to ensure higher yield and performance even under difficult conditions. Amplification can be detected in a number of ways including real-time fluorescence detection and end-point visualization. For added convenience the enzyme is available with separate fluorescent dye (enabling real-time detection with any qPCR thermocycler), and in a 2x mix format for those looking for reduced setup times. The enzyme gives consistent results regardless of the format chosen.