{"id":2092,"date":"2015-04-27T12:16:34","date_gmt":"2015-04-27T12:16:34","guid":{"rendered":"http:\/\/www.neb-online.de\/?page_id=2092"},"modified":"2024-07-17T12:37:58","modified_gmt":"2024-07-17T10:37:58","slug":"pcr-and-dna-amplification","status":"publish","type":"page","link":"https:\/\/www.neb-online.de\/en\/pcr-and-dna-amplification\/","title":{"rendered":"PCR and DNA Amplification"},"content":{"rendered":"

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The right DNA polymerase for your application<\/h2>\n

As the Polymerase Chain Reaction (PCR) is the most common DNA amplification method in molecular biology, NEB\u2019s product portfolio features a large selection of polymerases geared towards this powerful method. As the first company to sell Taq<\/em> DNA polymerase<\/a><\/span> to the research market and the first to discover a PCR-stable, high-fidelity DNA polymerase, NEB has a long history of developing reliable and convenient PCR tools.[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_tta_tabs style=”modern” spacing=”2″ active_section=”1″ css_animation=”none” no_fill_content_area=”true” el_class=”tab-menu-font-size”][vc_tta_section title=”More Information” tab_id=”1522220232076-52a08c14-0a933d6b-8fe12c28-5d22″][vc_column_text]<\/p>\n

OneTaq<\/em> DNA Polymerase & Q5 DNA Polymerase<\/h2>\n

NEBs commitment to innovative, high quality tools for DNA amplification and PCR products has continued with the development of OneTaq<\/em><\/a><\/span> for robust routine PCR and Q5 High-Fidelity DNA Polymerase<\/span><\/a> for robust, ultra high-fidelity PCR (~280x Taq<\/em> fidelity). Both product lines have been developed to tolerate a variety of complex templates without experiencing a loss of performance on high-AT or high-GC targets. A variety of NEB polymerases, including OneTaq<\/em>, Taq<\/em> and Q5, also benefit from novel aptamer-based hot start technology that does not require a separate activation step.[\/vc_column_text][vc_separator color=”black”][vc_column_text]<\/p>\n

Luna qPCR Products<\/h2>\n

Fluorescence-based quantitative real-time PCR (qPCR) is the gold standard for the detection and quantification of nucleic acids due to its sensitivity and specificity. Luna products from NEB<\/span><\/a> are optimized for qPCR or RT-qPCR, and are available for either intercalating dye or probe-based detection methods. All Luna products provide robust performance on diverse sample sources and target types.<\/p>\n

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Isothermal Amplification<\/h2>\n

Despite the ubiquitous nature of PCR, it may not be the best option for all amplification needs. For point of care and other diagnostic applications, sequence-specific isothermal amplification methods, that eliminate the need for thermocycling, have been particularly useful. Instead of heat, these methods typically employ a strand-displacing DNA polymerase, like Bst DNA Polymerase, Large Fragment<\/span><\/a>, to separate duplex DNA. To address some of the limitations of current isothermal amplification techniques, NEB has developed the next generation Bst, Bst<\/em> 3.0 , 2.0 und\u00a0\u00a0Bst<\/em> 2.0 WarmStart DNA Polymerasen<\/a><\/span> version of this enhanced polymerase, which enables room temperature reaction set up, yet is fully active at temperatures greater than 50\u00b0C.[\/vc_column_text][\/vc_tta_section][vc_tta_section title=”Videos” tab_id=”1522140363734-94ff0233-20b33d6b-8fe12c28-5d22″][vc_row_inner][vc_column_inner width=”1\/2″][vc_column_text]How to Choose the Right DNA Polymerase for PCR
\n<\/strong>Choosing the correct DNA polymerase for PCR depends on many factors. NEB offers a range of DNA polymerase formulations optimized for different applications. In this video Product Marketing Manager Fiona Stewart explains which products to choose for routine amplification, high-fidelity amplification, and next generation sequencing applications depending on the base content of your target amplicon. To choose the right DNA polymerase for your application please visit our PCR selection tool<\/a>
\n<\/span>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=”1\/2″][vc_video link=”https:\/\/www.youtube-nocookie.com\/embed\/z6vsdRaMOuE”][\/vc_column_inner][\/vc_row_inner][vc_separator color=”black”][vc_row_inner][vc_column_inner width=”1\/2″][vc_column_text]Important Tips for Working with Q5 Polymerase<\/strong>
\nAre you using Q5\u00ae High-Fidelity DNA Polymerase? Then you probably already know how robust and accurate the DNA amplification is (greater than 100X Taq DNA Polymerase). Melanie has some tips for ensuring optimal performance with Q5. If you haven’t tried Q5 for your high-fidelity amplification yet, what are you waiting for?<\/span>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=”1\/2″][vc_video link=”https:\/\/www.youtube.com\/watch?v=ity1fhL4uaM”][\/vc_column_inner][\/vc_row_inner][vc_separator color=”black”][vc_row_inner][vc_column_inner width=”1\/2″][vc_column_text]Why Choose Q5 High-fidelity Polymerase?
\n<\/strong>Q5\u00ae High-Fidelity DNA Polymerase sets a new standard for both fidelity and robust performance across the GC spectrum. With the highest fidelity amplification available (more than 100 times higher than Taq), Q5 DNA Polymerase results in ultra-low error rates. Q5 DNA Polymerase is composed of a novel polymerase that is fused to the processivity-enhancing Sso7d DNA binding domain, improving speed, fidelity and reliability of performance. In this video Senior Applications Scientist Nicole Nichols explains which Q5 formulation is optimal for your specific PCR application.<\/span>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=”1\/2″][vc_video link=”https:\/\/www.youtube-nocookie.com\/embed\/-UP5A08UOQE”][\/vc_column_inner][\/vc_row_inner][vc_separator color=”black”][vc_row_inner][vc_column_inner width=”1\/2″][vc_column_text]How to Amplify GC-rich DNA<\/strong>
\nAmplification of GC-rich DNA can be challenging with traditional DNA polymerases. NEB offers a range of DNA polymerase formulations optimized for the amplification of GC-Rich DNA. In this video Senior Applications Scientist Nicole Nichols explains which products to choose for routine amplification, high-fidelity amplification, and next generation sequencing applications depending on the base content of your target amplicon. <\/span>[\/vc_column_text][\/vc_column_inner][vc_column_inner width=”1\/2″][vc_video link=”https:\/\/www.youtube-nocookie.com\/embed\/wUjDZiruSLI”][\/vc_column_inner][\/vc_row_inner][vc_column_text]Mehr Video Tutorials finden Sie auf NEBs YouTube Kanal<\/a>.[\/vc_column_text][\/vc_tta_section][vc_tta_section title=”NEB Tools” tab_id=”tools”][vc_column_text]<\/p>\n

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NEB Tools<\/strong><\/h2>\n<\/div>\n<\/div>\n

[\/vc_column_text][vc_row_inner][vc_column_inner width=”1\/6″][vc_single_image image=”2015″ img_size=”full” onclick=”custom_link” img_link_target=”_blank” link=”https:\/\/tmcalculator.neb.com\/#!\/”][\/vc_column_inner][vc_column_inner width=”5\/6″][vc_column_text]Use this tool when designing PCR reaction protocols to help determine the optimal annealing temperature for your amplicon. Simply input your DNA polymerase, primer concentration and your primer sequence and the Tm Calculator<\/strong> will guide you to successful reaction conditions.[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=”1\/6″][vc_single_image image=”2017″ img_size=”full” onclick=”custom_link” img_link_target=”_blank” link=”http:\/\/nebiocalculator.neb.com\/#!\/”][\/vc_column_inner][vc_column_inner width=”5\/6″][vc_column_text]Use this tool for your scientific calculations and conversions for DNA and RNA. Options include conversion of mass to moles, ligation amounts, conversion of OD to concentration, dilution and molarity. Additional features include sgRNA Template Oligo Design and qPCR library quantification.[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=”1\/6″][vc_single_image image=”2016″ img_size=”full” onclick=”custom_link” img_link_target=”_blank” link=”https:\/\/nebasechanger.neb.com\/”][\/vc_column_inner][vc_column_inner width=”5\/6″][vc_column_text]NEBaseChanger<\/strong> can be used to design primers specific to the mutagenesis experiment you are performing using the Q5 Site-Directed Mutagenesis Kit. This tool will also calculate a recommended custom annealing temperature based on the sequence of the primers by taking into account any mismatches.[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=”1\/6″][vc_single_image image=”2011″ img_size=”full” onclick=”custom_link” img_link_target=”_blank” link=”http:\/\/pcrselector.neb.com\/#!\/#%2F”][\/vc_column_inner][vc_column_inner width=”5\/6″][vc_column_text]Use this tool to help select the right DNA polymerase for your PCR setup. Whether your amplicon is long, complex, GC-rich or present in a single copy, the PCR selection tool<\/strong> will identify the perfect DNA polymerase for your reaction.[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=”1\/6″][vc_single_image image=”14067″ img_size=”62×64″ onclick=”custom_link” img_link_target=”_blank” link=”http:\/\/pcrfidelityestimator.neb.com\/#!\/”][\/vc_column_inner][vc_column_inner width=”5\/6″][vc_column_text]Estimate the percentage of correct DNA copies (those without base substitution errors) per cycle of PCR for selected DNA polymerases.[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=”1\/6″][vc_single_image image=”9562″ img_size=”full” onclick=”custom_link” img_link_target=”_blank” link=”https:\/\/www.neb.com\/en\/tools-and-resources\/selection-charts\/dna-polymerase-selection-chart”][\/vc_column_inner][vc_column_inner width=”5\/6″][vc_column_text]This table lists properties that should be considered when choosing a polymerase. Since these properties can depend on reaction conditions, the primary references should be consulted prior to use in a given application.[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_tta_section][vc_tta_section title=”Guides” tab_id=”1721043910706-156f3779-9550″][vc_column_text]<\/p>\n

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Troubleshooting Guides<\/h3>\n