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DTI JadeAmp FabTaq Premix PCR Master Mix is a loading-dye-added version of JadeAmp FabTaq MAX PCR Master Mix that is optimized for great performance and convenience in both standard and high-throughput PCR applications. This master mix includes an optimized buffer, PCR enzyme, dNTP mixture, gel loading dye (green), and a density reagent in a 2X premix format. Simply add primers and DNA template. Following PCR, amplicons can be used directly in several ways.

PCR tube contents can be loaded directly onto an agarose gel for electrophoresis or used directly in downstream applications such as restriction enzyme digestion, TA cloning, and direct sequencing. DTI JadeAmp FabTaq Premix PCR Master Mix can be used to amplify genomic targets up to ~5 kb and is compatible with GC- and AT-rich targets.

When 5 µl of the loading-dye-added PCR master mix is used for electrophoresis on an 1% Agarose gel, the blue dye front is detected around 3–5 kb, and the yellow dye is detected below 50 bp. These dyes have absorptions at approximately 260 nm and 420 nm, respectively. The dyes can be removed by gel purification or PCR cleanup using the NucleoSpin Gel and PCR Clean-Up kit (where available), if necessary.

Specifications

Documents

Applications

FAQs

Brand	Catalogue Number	Pack Size
DTI JadeAmp FabTaq Premix	DT0201.80	80 rxn (25 μl volume)
DTI JadeAmp FabTaq Premix	DT0201.320	320 rxn (25 μl volume)

• Convenient premix format—just add template, primers, and water to the 2X PCR master mix.

• Improved performance—insludes an optimized buffer for better amplification of GC- and AT-rich targets.

• Eliminates purification steps—PCR contents can be loaded directly onto an agarose gel.

• High-throughput PCR—use reactions directly in TA cloning, restriction enzyme digestion, or sequencing.

• Green loading dye—allows tracking during gel electrophoresis.

DTI JadeAmp FabTaq Premix - Protocol

DTI JadeAmp FabTaq Premix - Certificate of Analysis

DTI JadeAmp FabTaq Premix - Promotional Document

DTI JadeAmp premixes outperforms Competitor(C) Red mix for GC-rich targets

Results

Amplification of GC-rich targets

Our results showed that the non-hot-start DTI JadeAmp FabTaq Premix and DTI JadeAmp Max HiFid Taq Premix gave minimal background amplification with GC-rich targets. Non-hot-start Competitor(C) Red Mix, on the other hand, resulted in nonspecific amplification and was unable to amplify the target with a high GC content (72.3%). While DTI JadeAmp FabTaq Premix failed to amplify the TGFB-1 gene that is 4 kb long with a 63.1% GC content, DTI JadeAmp Max HiFid Taq Premix was able to generate a clean and distinct band. We were unable to amplify the 4-kb TGFB-1 target using Competitor(C) Red Mix.

Figure 1: Comparison of amplification profiles of four GC-rich target genes using DTI JadeAmp FabTaq Premix, DTI JadeAmp Max HiFid Taq Premix, or Competitor(C) Red Mix. 10 µl from each PCR reaction was loaded on a 1% agarose gel.

Conclusions

DTI JadeAmp FabTaq Premix and DTI JadeAmp Max HiFid Taq Premix (non-hot-start and hot-start) outperformed Competitor(C) Red Mix in background amplification, target GC-content, and target length. The DTI JadeAmp series gave minimal background amplification and better performance on GC-rich targets. Additionally, the DTI JadeAmp Max HiFid Taq Premix master mixes were able to amplify long targets while Competitor(C) Red Mix failed to amplify the 4-kb target gene TGFB-1.

With DTI's JadeAmp series of PCR master mixes, you can streamline your genotyping workflow without worrying about GC content of your target or nonspecific background amplification.

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.

What is the optimal amount of DNA template that should be used for PCR?

The optimal amount of template required depends on the complexity of the template and the copy number of the target sequence. Approximately 104 copies of the target DNA sequence are required to detect the amplification product in 25–30 PCR cycles.

• Typically, 1 µg of human genomic DNA contains 3.04 x 105 molecules of DNA. For most PCR applications, 30–100 ng of human genomic DNA is sufficient. High-copy targets, such as housekeeping genes, require only 10 ng of template. Template amounts for higher-complexity templates range between 10 ng and 500 ng.

• Typically, 1 µg of E. coli genomic DNA contains 2 x 108 molecules of DNA; therefore, the recommended amount of template is between 100 pg and 1 ng.

• Typically, 1 µg of lambda DNA contains 1.9 x 1010 molecules of DNA; therefore, the template input can be as little as 100 pg.

• The amount of cDNA template depends on the copy number of the target. cDNA input is typically described in terms of equivalent RNA input. The amount of cDNA in a PCR reaction can be as little as 10 pg (RNA equivalent).

For some Polymerase, excessive template tends to inhibit amplifications. In contrast, some DNA Polymerase tolerates a broader range of template quantity. In general, reaction conditions may be adjusted depending on the type, quantity, and quality of template.

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