Reduced reaction volumes and increased Taq DNA polymerase concentration improve STR profiling outcomes from a real-world low template DNA source: telogen hairs.
OBJECTIVE The main method for the analysis of low template DNA (LTDNA) is known as a method of low copy number (LCN), which involves increasing the number of PCR cycles (typically 34). In common with other LTDNA method, characterized by an imbalance LCN profile allele, decline, and the drop outs which require interpretation of complex rules. They often require replicate PCR reactions to produce a “consensus” profile in special facilities. The ideal method for the analysis of LTDNA should raise the profile of the results without a high error rate and is accomplished using the standard amenities, in addition to the minimum cost.
METHOD In this study, we present a comparison of four methods of variation for amplification of STR from LTDNA broadly used, the kit is available commercially (AmpFℓSTR (®) Profiler Plus (®)): the standard method, the standard method to post a PCR clean up, the method of the LCN, and the reaction volume decreases with increased concentrations of Taq DNA polymerase.
RESULTS Using the telogen hair-LTDNA common source and reference DNA match, LCN method produces the highest number of appropriate and non-appropriate alleles (ie, down-in). For comparison, the reaction volume was reduced by an increase in Taq Polymerase produce fuller profile and corresponding DNA (all alleles combined) and less off-ladder alleles of a variety of input DNA. In addition, this method results in less non-concordant allele of the LCN and not more than for standard PCR, which indicates that it may be preferable to increase PCR cycles for analysis LTDNA, either with or without consensus profiles and statistical modeling.
CONCLUSION Overall, this study highlights the importance and benefits of optimizing PCR conditions and to develop improved laboratory method to amplify and analyze LTDNA.
Reduced reaction volumes and increased Taq DNA polymerase concentration improve STR profiling outcomes from a real-world low template DNA source: telogen hairs.
[Effect of Incorporation Efficiency Charge Chromophore on fluorescently-labeled nucleotides in Matrix synthesis by Taq DNA Polymerase].
In order to study the effect of the electric charge of the chromophore, the efficiency of incorporation of fluorescently labeled nucleotides into DNA during PCR, three fluorescently labeled dUPT, one with dye electroneutral and the other two with positive and negative (Cy5 analog), was synthesized.
This shows that dUPT, labeled with Cy 5 electroneutral analog, most effectively incorporated into the DNA when Tag polymerase used for PCR. Polymerase chain reaction (PCR) is widely used in a variety of experimental conditions, and Taq DNA polymerase is crucial in the process of PCR. In this article, Taq DNA expression polymerase plasmid reconstructed and protein products obtained by purification fast, ( “Fast purification of high-activity of Taq DNA polymerase” (Pluthero, 1993 [1]), “Single-step purification of DNA thermostable polymerase expressed in Escherichia coli “(Desai and Pfaffle, 1995 [2])). Here we present data on production of protein expression and provides the analysis result of the production of two different vectors. Meanwhile, refining the data is also provided to indicate the purity of the protein product.
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer10mM dNTP Mix
Description: Taq DNA Polymerase is a highly thermostable DNA Polymerase that catalyzes the 5’-3’ synthesis of DNA. This polymerase has 5’-3’ exonuclease activities, lacks 3’-5’ exonuclease activity, and produces 3’-dA-tailed amplicons. PCR products made with Taq can be used with TA cloning vectors.
Description: The enzyme is supplemented with two inert gel tracking dyes (red and yellow),allowing direct loading of PCR products to agarose gels (1.0 U/μl)
Description: The enzyme is supplemented with two inert gel tracking dyes (red and yellow),allowing direct loading of PCR products to agarose gels (1.0 U/μl)
Description: The enzyme is supplemented with two inert gel tracking dyes (red and yellow),allowing direct loading of PCR products to agarose gels (1.0 U/μl)
Description: The enzyme is supplemented with two inert gel tracking dyes (red and yellow),allowing direct loading of PCR products to agarose gels (1.0 U/μl)
Description: Intact Genomics Taq DNA Polymerase 2x Premix with Dye is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends. Intact Genomics Taq DNA Polymerase 2x Premix with Dye is ready to use, containing Taq DNA Polymerase, dNTPs, MgCl2, and stabilizers. It has been optimized for routine PCR applications.Product Includes:Taq DNA Polymerase 2x Premix with Dye5x Magic Enhancer
Description: Intact Genomics Taq DNA Polymerase 2x Premix with Dye is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends. Intact Genomics Taq DNA Polymerase 2x Premix with Dye is ready to use, containing Taq DNA Polymerase, dNTPs, MgCl2, and stabilizers. It has been optimized for routine PCR applications.Product Includes:Taq DNA Polymerase 2x Premix with Dye5x Magic Enhancer
chemical modifications to DNA, such as modification 2 ‘, is expected to increase the utility of DNA biotechnology; However, other forms of modification of DNA is limited by their inability to effectively synthesized by a DNA polymerase enzyme. Previous efforts have identified a mutant Thermus aquaticus DNA polymerase I (Taq) enzyme that can recognize DNA 2’-modified nucleotides. While these modified nucleotides recognize the mutant enzyme, they are unable to synthesize the full length of the modified DNA; thus, further techniques necessary for this enzyme
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