Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) is formulated without glycerol to support lyophilization, incorporation into microfluidic devices, and enable quick adoption into automation workflows.
Bst 2.0 WarmStart DNA Polymerase is an in silico designed homolog of Bacillus stearothermophilus DNA Polymerase I, Large Fragment (Bst DNA Polymerase, Large Fragment) with a reversibly-bound aptamer, which inhibits polymerase activity at temperatures below 45°C. At temperatures above 45°C, the aptamer rapidly releases from the enzyme with no special activation step needed to activate the polymerase. Bst 2.0 WarmStart DNA Polymerase contains 5´→3´ DNA polymerase activity and strong strand-displacement activity but lacks 5´→3´ exonuclease activity. Bst 2.0 WarmStart DNA Polymerase displays improved amplification speed, yield, salt tolerance, and thermostability compared to wild-type Bst DNA Polymerase, Large Fragment.
Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) offers the same robust detection of human DNA targets in LAMP assays as the glycerol-containing enzyme.
LAMP (DNA targets) experiments were performed with NEB #M0538: Bst 2.0 WarmStart DNA Polymerase or NEB #M0402: Bst 2.0 WarmStart DNA Polymerase (Glycerol-free). Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent dye were set up in quadruplicate over three logs of total Jurkat DNA (10 ng to 0.1 ng) in 96-well, 25 µl reactions. Control reactions without template (NTC) were also evaluated. Reactions were incubated at 65°C for 40 minutes and fluorescence was monitored every 15 seconds in the SYBR/FAM channel of a real-time thermocycler (Bio-Rad® CFX96). Each dot represents the time at which the fluorescence signal for a single reaction crosses the instrument-defined threshold. All four replicates were detected at each template input unless otherwise indicated (note that dots frequently overlap given similar detection time for the replicates). Overall, similar performance was observed for both glycerol-containing (NEB #M0538) and glycerol-free (NEB #M0402) enzymes at each template input. No amplification was observed in any of the no template control reactions.
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