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, a 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 the need for further purification.

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.

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.

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.

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

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