DTI FabTaq HS-Glycerol Free (50 U/ul) is a hot start PCR enzyme derived from Thermus aquaticus that includes a neutralizing monoclonal antibody that recognizes Taq DNA polymerase. This antibody binds to Taq DNA polymerase and prevents non-specific amplification due to mispriming and/or formation of primer dimers before thermal cycling. The antibody is denatured during the initial DNA-denaturation step, allowing this product to be used with standard PCR conditions. Since the product is glycerol free, it is suitable for lyophilization processes
| Catalog # | Pack Size |
|---|---|
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DT0103.2500 |
2500 U
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DTI CobaltAmp HiFid Taq HS Premix is a Fast PCR Master Mix and designed for streamlined PCR, and is, therefore, ideal for applications such as colony PCR. It contains a hot-start PCR enzyme, optimized buffer, dNTPs, along with a blue gel-loading dye and density reagent, in a convenient 2X premix format. Reactions are assembled by simply mixing primers and DNA template with DTI CobaltAmp HiFid Taq HS Premix. For rapid and convenient screening, the amplification reactions can be loaded directly onto an agarose gel immediately after amplification without purification, or digested directly with restriction enzymes.
Storage and Shipment Conditions
-80°C
Components
| Components | Quantity |
|---|---|
| DTI FabTaq HS-Glycerol Free (50 U/μl) | 50 µl |
| * Customized pack size available |
Materials required but not provided:
10X PCR buffer
25 mM/50 mM MgCl2
2.5 mM dNTP
Template DNA/cDNA
Target specific forward and reverse primers
Nuclease free water
Thermal cycler
Agarose electrophoresis system
Loading dyes and marker
Unit Definition
One unit is the amount of enzyme that will incorporate 10 nmol of dNTPs into acid-insoluble products in 30 minutes at 74°C with activated salmon sperm DNA as the template-primer.
Application
For DNA amplification by hot start PCR
For DNA sequencing
For further manufacturing and lyophilization processes
Reaction mixture for unit definition :
| 25 mM TAPS (pH 9.3 at 25℃) | 50 mM KCl |
| 2 mM MgCl2 | 0.1 mM DTT |
| 200 μM each dATP·dGTP·dCTP | 100 μM [3H]-dTTP |
| 0.25 mg/ml activated salmon sperm DNA |
Purity
Nicking, endonuclease, and exonuclease activity were not detected after incubation of 0.6 μg of supercoiled pBR322 DNA, 0.6 μg of λDNA, or 0.6 μg of λ-Hin d III digest with 10 U of this enzyme for 1 hour at 74°C.
Quality Control Data
Please see the certificate of analysis for each lot.
| Document | Download Link |
|---|---|
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DTI FabTaq HS-Glycerol Free (50U/ul) – Protocol |
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DTI FabTaq HS-Glycerol Free (50U/ul) – Certificate of Analysis |
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DTI FabTaq HS-Glycerol Free (50U/ul) – Promotional Document |
Experimental Sample
a) Amplification data: Good performance of this product has been confirmed by PCR using λ DNA as template (amplified fragment size 8 kbp). The protocol used for the assay and the results are as follows:
Figure 1: Amplification data showing product performance with λ DNA as template.
b) Hot start function data: Test target sample added to a PCR system containing FabTaq and FabTaq HS respectively. Following an incubation at RT for 30 mins, the products were amplified using a thermal cycler. Non-specific bands (around 280 bp) were observed in the PCR system along with specific target bands (838 bp) using FabTaq, while there were no non-specific bands observed in PCR system using FabTaq HS.
Figure 2: Amplification data for λ DNA (8 kbp amplified fragment size).
Figure 3: Additional data showcasing product performance.
Figure 4: Hot start function data showing non-specific bands in PCR system with FabTaq and absence of non-specific bands with FabTaq HS.