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Bst 2.0 WarmStart DNA Polymerase is an in silico designed homologue 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. The aptamer rapidly releases the Bst 2.0 WarmStart DNA Polymerase above 45°C and therefore no special activation step is 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.

Benefits of Bst 2.0 WarmStart
Identical LAMP reactions were run either immediately after setup (solid line) or after a 2 hour incubation at 25 °C. Without the protection from Bst 2.0 WarmStart, this room temperature incubation results in variable LAMP performance. Bst 2.0 WarmStart provides more consistent amplification reaction and enables room-temperature and high-throughput setup.

产品来源

Bst 2.0 WarmStart DNA Polymerase is prepared from an E. coli strain that expresses the Bst 2.0 DNA Polymerase protein from an inducible promoter.

应用特性

• Isothermal amplification (LAMP)

• Applications requiring strand-displacement DNA synthesis

• DNA sequencing through high GC regions

• Rapid sequencing from nanogram amounts of DNA template

单位定义

One unit is defined as the amount of enzyme that will incorporate 25 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.

反应条件

1X Isothermal Amplification Buffer Pack
Incubate at 65°C

1X Isothermal Amplification Buffer Pack
20 mM Tris-HCl
10 mM (NH4)2SO4
50 mM KCl
2 mM MgSO4
0.1% Tween® 20
(pH 8.8 @ 25°C)

贮存溶液

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% Triton® X-100
pH 7.1 @ 25°C

热失活

80°C for 20 min

单位活性检测条件

50 mM KCl, 20 mM Tris- HCl (pH 8.8), 10 mM MgCl2, 30 nM M13mp18 SS DNA, 70 nM M13 sequencing primer (–47) 24 mer, 200 μM dATP, 200 μM dCTP, 200 μM dGTP, 100 μM dTTP including [³H]-dTTP and 100 μg/ml BSA.

注意事项

1. Bst 2.0 WarmStart DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.

2. Reaction temperatures above 70°C are not recommended.

3. Bst 2.0 WarmStart DNA Polymerase cannot be used for thermal cycle sequencing or PCR.

4. Specific reaction conditions will vary for differentisothermal amplification applications. For best results, use 1X Isothermal Amplification Buffer.