Low DNA Taq HS 5 U/µL
Low DNA Taq HS, 5 U/µL is a highly purified chemically modified hot start Taq DNA polymerase and buffer system, with separate magnesium. It has been developed for low bioburden and very high sensitivity over a wide range of PCR templates.
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High sensitivity and specificity using Low DNA Taq HS
A 10-fold dilution of DNA was used in a qPCR assay using Low DNA Taq HS and an intercalating dye. Each dilution was run in triplicate using standard reaction conditions. A/ The amplification curve demonstrates the reproducibility and sensitivity of the Low DNA Taq HS and B/ The single distinct peak in the melt curve illustrates the products are a single discrete species, with no additional bands or primer/dimers.
Low DNA Taq HS, 5 U/µL, MDX009
Available in 0.1 mL (500 Units) or 10 mL (50,000 Units) aliquots
Description
A chemical moiety is attached to Low DNA Taq HS, 5 U/µL making it inactive at ambient temperature, avoiding extension of non-specifically annealed primers or primer dimers and providing higher specificity of DNA amplification. Low DNA Taq HS, 5 U/µL catalyzes 5′-3′ synthesis and possesses low 5′-3′ exonuclease activity, however these are not detectable before activation, enhancing PCR sensitivity. The functional activity of the enzyme is restored during a 10-minute incubation at 95 °C. The low DNA background and stringent hot-start properties of Low DNA Taq HS, 5 U/µL are ideal for PCR of low-copy bacterial targets and avoiding false-positive amplification, such as in water testing.
Specifications
| Description | A highly purified Taq DNA polymerase. The low DNA background and stringent hot-start properties are ideal for PCR of low-copy bacterial targets and avoiding false-positive amplification, such as in water testing. |
| Concentration | 5 U/µL |
| Appearance | Clear, colorless solution |
| Hot Start | Chemical |
| Purity | >90% |
| Application | PCR, two-step RT-PCR |
| Presentation | 1 vial |
| Storage | -20 °C |
| Mix stability | See outer label |
| Consistency | ±0.5 Ct variance between test and reference sample |
| DNA Contamination | None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. |
| DNase Contamination | No detectable degradation |
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FAQs: Low DNA qPCR mix
With a chemical hot start Taq polymerase, the polymerase is covalently linked with chemical groups to block enzyme activity at room temperature.
Advantages
• More stringent than antibody hot-start methods, so there is no activity even for long periods at room temperature, minimizes non-specific amplification
• Low bioburden, as it is free of animal and bacterial-origin components
Considerations
• Longer activation time required for the polymerase to become fully active
Low DNA Taq HS, 5 U/µL lacks a 3′ – 5′ exonuclease activity. However, the enzyme does have 5 ‘ – 3′ nuclease activity. During the extension step of a PCR amplification, the enzyme will hydrolyze any blocking strand starting from its 5’ end.
Low DNA Taq HS, 5 U/µL lacks proofreading activity, so it will leave a 3′-overhang which can be used with TA cloning. In order to drive the reaction to the extra A state, a final extension time at 72°C should be increased to 15-30 minutes.
Yes, once activated, Low DNA Taq HS, 5 U/µL remains active. Lowering the temperature will not inactivate Low DNA Taq HS, 5 U/µL.
No the 10-minute activation will the enzyme, it will remain fully functional during the subsequent cycling.
Multiplex PCR involves the co-amplification of multiple amplicons in a single PCR. Since several sets of primers are being added to a single reaction, the potential for primer dimer formation or mispriming and so loss of specificity and so a decrease in yield of specific product exists. Because Low DNA Taq HS, 5 U/µL remains active has no activity prior to the hot start PCR, the multiple primers do not have the possibility to react with themselves, dramatically increasing specific product yield.
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