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DTI FabTaq HS DNA Polymerase is the hot-start version of our high-performing DTI FabTaq polymerase offering high yield, excellent sensitivity, and fidelity that is 4.5 times higher than Taq polymerase. Antibody-mediated hot-start gives lower background, higher specificity, and allows room temperature reaction assembly. DTI FabTaq HS DNA polymerase is supplied with 10X buffer (Mg2+) and dNTP mix.

Specifications

Documents

Applications

FAQs

Brand	Catalogue Number	Pack Size
DTI FabTaq HS	DT0102.250	250 U

• Antibody-mediated hot start: lower background, increased specificity, and room-temperature reaction assembly.

• High-yield and high-sensitivity PCR.

• Excellent on difficult templates.

DTI FabTaq HS - Protocol

DTI FabTaq HS - Certificate of Analysis

DTI FabTaq HS - Promotional Document

Results

Comparison with a DTI FabTaq HS DNA polymerase

The performance of DTI FabTaq HS in a multiplex reaction was compared to its performance in single-target reactions, and to that of the standard DTI FabTaq DNA Polymerase in a multiplex reaction (Figure 1). DTI FabTaq and DTI FabTaq HS were each used to amplify a human genomic DNA template with eight different primer pairs, each specific for a target ranging from 84 to 432 bp in size. In addition, DTI FabTaq HS was used to perform an individual amplification reaction with each of the eight primer pairs.

The sample amplified using DTI FabTaq HS (Lane 10) shows more efficient multiplex amplification of all the bands individually amplified in Lanes 1–8 than the sample amplified using DTI FabTaq (Lane 9), and it lacks the low molecular weight non-specific amplification band seen with DTI FabTaq (Lane 9). The intensities of the bands in Lane 10 are comparable to the intensities of the corresponding individual bands in Lanes 1–8, indicating that DTI FabTaq HS provides multiplex efficiencies comparable to those observed in individual amplification reactions.

Comparing the ability of DTI FabTaq DNA Polymerase Hot Start Version to amplify human genomic DNA fragments to that of the standard DTI FabTaq DNA Polymerase. PCR reactions were performed using human genomic DNA as a template and primer pairs for eight different targets. Lanes 1–8 contain individual reactions for each primer pair amplified using DTI FabTaq HS. Lanes 9 and 10 contain multiplex PCR reactions performed with all eight primer pairs in a single tube, amplified with either the standard Taq DNA polymerase (DTI FabTaq, Lane 9) or DTI FabTaq HS (Lane 10).

Conclusions

This experiment demonstrates that target amplification efficiencies for multiplex PCR using DTI FabTaq HS are comparable to efficiencies observed for separate (single target) amplification reactions. In addition, DTI FabTaq HS demonstrates superior efficiency and specificity over standard Taq polymerase (DTI FabTaq) in this multiplex-PCR application.

Methods

PCR cycling conditions for individual reactions

1 cycle at 94°C for 30 sec, followed by 30 cycles of 94°C for 30 sec, 55°C for 30 sec, and 72°C for 60 sec.

PCR cycling conditions for multiplex reactions

1 cycle at 94°C for 30 sec, followed by 30 cycles of 94°C for 30 sec, 57°C for 30 sec, and 72°C for 60 sec, followed by a final step at 72°C for 90 sec.

Which enzymes faciliate efficient workflow when analyzing multiple samples by gel electrophoresis after PCR (e.g., genotyping screens)?

We recommend DTI JadeAmp FabTaq Premix (Cat.# DT0201.320). This product is completely premixed, making it easy to prepare PCR reaction mixtures which can be loaded directly on an agarose gel for electrophoresis after the reaction. For an efficient workflow with minimal pipetting, DTI JadeAmp FabTaq Premix contains a green tracking dye. Additionally, density agent is included to aid in gel loading.

DTI JadeAmp FabTaq Premix can amplify targets up to 10 kb in length, including targets that are GC- or AT-rich. Additionally, PCR products generated with DTI JadeAmp FabTaq Premix can be used directly for restriction enzyme digestion, sequencing, or TA-cloning without need for further purification.

Which enzymes are suitable for colony PCR?

We recommend DTI CobaltAmp HiFid Taq HS Premix for colony PCR. These enzyme preparations tolerate the presence of substantial bacterial nucleic acid carry-over. Because tracking dye and density agent are included in these master mixes, the PCR reaction mixtures may be loaded directly on an agarose gel for electrophoresis.

Which enzymes are suitable for fast PCR?

DTI CobaltAmp HiFid Taq HS Premix is an economical choice for high-throughput projects. This enzyme is formulated to include high speed polymerase, optimized buffer, dNTP mixture, gel loading dye (blue), and a density reagent in a 2X premix. Since it requires an extension time of only 10 sec/kb, colony PCR reactions can be completed in less than 1 hour for inserts up to 1 kb.

What precautions should be taken when using an inosine-containing primer?

DTI FabTaq and DTI FabTaq Hot Start Version enzymes are compatible with inosine-containing primers. It is important to note that inosine-containing primers should not be used with PCR enzymes that have 3’→5’ exonuclease activity. When using one of these PCR enzymes, we recommend using mixtures of degenerate primers with A, T, G, and C at the desired position(s) rather than inosine-containing primers when performing degenerate PCR.

What is the terminal structure of DTI FabTaq and DTI FabTaq Hot Start Version enzymes’ amplification product?

DTI FabTaq and DTI FabTaq Hot Start Version enzymes primarily yield amplification products containing 3’-dA overhangs, which facilitates TA cloning.

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