Request a hot-start sample Why use green? The two tracking dyes and a density reagent in the green buffer do not interfere with PCR performance, and are compatible with downstream applications including DNA sequencing, ligation, and restriction digestion. This hot-start DNA polymerase offers higher yields and longer amplicons than conventional Taq-based products. In addition, due to the hot-start modification, DreamTaq Hot Start DNA Polymerase has been engineered to provide increased sensitivity and specificity. In addition, the green buffer format provides the convenience of direct loading of PCR products on a gel.
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Green buffer is a proprietary formulation optimized for robust performance in PCR. The dyes have absorption peaks at nm and nm. Therefore, care must be taken to avoid contamination with other templates and amplicons that may be present in the laboratory environment. General recommendations to lower the risk of contamination are as follows:? Wear fresh gloves for DNA purification and reaction set up. Use reagent containers dedicated for PCR. Use positive displacement pipettes, or pipette tips with aerosol filters to prepare DNA samples and perform PCR set up.
PCR primers are generally nucleotides long. Ideally, C and G nucleotides should be distributed uniformly along the primer. Avoid self-complementary primer regions, complementarities between the primers and direct primer repeats to prevent hairpin formation and primer dimerization.
Check for possible sites of undesired complementary between primers and template DNA. Estimation of primer melting temperature For primers containing less than 25 nucleotides, the approx. Higher amounts of template increase the risk of generation of non-specific PCR products. Lower amounts of template reduce the accuracy of the amplification. All routine DNA purification methods can be used to prepare the template e. Primers The recommended concentration range of the PCR primers is 0.
Excessive primer concentrations increase the probability of mispriming and generation of non-specific PCR products. For degenerate primers and primers used for long PCR we recommend higher primer concentrations in the range of 0.
Ordering Information Component DreamTaq? Gently vortex and briefly centrifuge DreamTaq? Place a thin-walled PCR tube on ice and add the following components for each 50? DNA polymerase, optimized DreamTaq? The master mix is supplemented with two tracking dyes and a density reagent that allows for direct loading of the PCR product on a gel. The dyes in the master mix do not interfere with PCR performance and are compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.
The master mix retains all features of DreamTaq? DNA polymerase. For applications that require PCR product analysis by absorbance or fluorescence excitation, we recommend using the colorless DreamTaq? Gently vortex the samples and spin down. When using a thermal cycler that does not contain a heated lid, overlay the reaction mixture with 25?
Load ? Annealing for 30 seconds is normally sufficient. Extension The optimal extension temperature for DreamTaq?
Number of cycles If less than 10 copies of the template are present in the reaction, about 40 cycles are required. For higher template amounts, cycles are sufficient. Troubleshooting For troubleshooting please visit www. Ribonuclease Assay Less than 0. Functional Assay DreamTaq? Notice to purchaser? In certain countries use of this product is covered by patents. For more information please contact info fermentas.
This product is licensed under one or more U. Patents Nos. This product is licensed under U. Patent No.
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