Bst-XT WarmStart™ DNA Polymerase (Glycerol-free) is formulated without glycerol to support lyophilization, incorporation into microfluidic devices, and quick adoption into automation workflows.
Bst-XT WarmStart DNA Polymerase is an engineered variant of Bacillus stearothermophilus DNA Polymerase, Large Fragment fused to a novel nucleic acid binding domain for improved isothermal amplification performance. This enzyme combines the high specificity of Bst 2.0 and the fast polymerization speed of Bst 3.0 DNA polymerases.
Bst-XT WarmStart DNA Polymerase contains a reversibly bound aptamer, which inhibits polymerase activity at room temperature. The aptamer rapidly releases above 45°C, therefore no separate activation step is needed.
Bst-XT WarmStart DNA Polymerase contains 5´→3´ DNA polymerase activity with either DNA or RNA templates and strong strand displacement activity, but lacks 5´→3´ and 3´→5´ exonuclease activity.
Bst-XT WarmStart™ DNA Polymerase (Glycerol-free) has equivalent performance to glycerol-containing Bst-XT WarmStart DNA Polymerase
LAMP (DNA targets, left panel) or RT-LAMP (RNA targets, right panel) experiments were performed with Bst-XT WarmStart DNA Polymerase (NEB #M9204) and Bst-XT WarmStart DNA Polymerase (Glycerol-free) (NEB #M9205) in Bst-XT isothermal amplification buffer. To detect RNA targets, WarmStart® RTx Reverse Transcriptase (NEB #M0380) was included in testing. Reactions containing 1X LAMP primers and 0.5X LAMP Fluorescent Dye (NEB #B1700) were set up in triplicate over three logs of total human Jurkat total RNA or Jurkat gDNA (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 30 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 three 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 and glycerol-free enzymes at each template input.
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Amplification Speed
Specificity
Room temp. set-up?
Available glycerol-free
