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 |
|---|---|
|
DT0103.2500 |
2500 U
|
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
| 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 |
|---|---|
|
DTI FabTaq HS-Glycerol Free (50U/ul) – Protocol |
|
|
DTI FabTaq HS-Glycerol Free (50U/ul) – Certificate of Analysis |
|
|
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.