Reduces non-specific product formation in isothermal amplification (e.g. LAMP)
This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.
Tte UvrD Helicase is a repair helicase capable of unwinding double-stranded DNA, without a requirement for a specific flap or overhang structure, from the thermophilic organism Thermoanaerobacter tengcongensis. It is active on a wide range of DNA substrates and, along with its thermostability (active to 70°C), Tte UvrD Helicase has been demonstrated to be a useful additive for improving specificity of isothermal amplification reactions, particularly in conjunction with the WarmStart® LAMP Kit (DNA & RNA).
Tte UvrD Helicase Eliminates Non-Template Amplification in LAMP Reactions
LAMP reactions were performed using the WarmStart® LAMP Kit (DNA & RNA) (NEB #E1700) and a primer set prone to high levels of non-template amplification. Without Tte UvrD Helicase (left), the positive reaction (+DNA) amplifies in ~8 minutes while the no-template control (NTC) amplifies in ~20 minutes. In the presence of 10 ng Tte UvrD Helicase (right), the positive reaction maintains its rapid amplification time with a slight reduction in total RFU, while the NTC reaction is completely suppressed.
Product Source
Tte UvrD Helicase is isolated from a strain of E. coli which carries the cloned UvrD gene from Thermoanaerobacter tengcongensis.
This product is related to the following categories:
1X Isothermal Amplification Buffer Pack
Supplement with 1 mM ATP
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)
Usage Concentration
0.2 - 0.4 ng/μl
Storage Buffer
10 mM Tris-HCl
100 mM KCl
1 mM DTT
0.1 mM EDTA
0.1% Triton® X-100
50% Glycerol
pH 7.4 @ 25°C
Heat Inactivation
80°C for 5 minutes
References
An, L., Tang, W., Ranalli, T.A., Kim, H.J., Wytiaz, J., and Kong, H. (2005). Characterization of a thermostable UvrD helicase and its participation in helicase-dependent amplification. J Biol Chem. 280 (32), 28952-8. PubMedID: 15955821
Mauris, J., and Evans, T.C. Jr. (2010). A human PMS2 homologue from Aquifex aeolicus stimulates an ATP-dependent DNA helicase. J Biol Chem. 285 (15), 11087-92. PubMedID: 20129926
Vincent, M., Xu, Y., and Kong, H. (2004). Helicase-dependent isothermal DNA amplification. EMBO Rep. 5 (8), 795-800. PubMedID: 15247927
Zaczek-Moczydłowska, M.A.; Mohamed-Smith, L.; Toldrà, A.; Hooper, C.; Campàs, M.; Furones, M.D.; Bean, T.P.; Campbell, K. (2020) A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1. Int J Mol Sci; 21, 6605,
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