DNA Amplification

Specialty PCR

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  • Some PCR experiments require polymerases optimized for specific applications. These reactions may include long or challenging templates that standard DNA polymerases don’t amplify well, reaction conditions that include suboptimal components or even inhibitors, or application-specific requirements.

    Long Range PCR: Optimized polymerase formulations can enable amplification of templates up to 30 kb and beyond. For Long Range PCR NEB provides LongAmp® and Q5® DNA polymerases. LongAmp is capable of amplifying very large fragments (up to 30 kb) with 2X Taq fidelity. Q5 can amplify fragments up to 10 kb, at the highest fidelity currently available on the market (> 100X Taq).

    Bisulfite Sequencing: Bisulfite-converted DNA used in epigenetic analysis contains uracil. Use of polymerases like EpiMark® Hot Start Taq (M0490) that have been optimized to amplify bisulfite-converted and AT-rich amplicons ensures robust amplification.

    1. Types of PCR

      NEB offers a variety of DNA polymerases for all of your experimental needs. High-fidelity, multiplex, and fast PCR are just a few of the types of amplification reactions you may need to conduct, and Breton explains which polymerases to choose.

    Extraction-Free/Direct PCR: Extraction-Free or Direct PCR describes amplification from blood, soil or other complex samples without an initial purification step. The presence of inhibitors in these sample types can reduce successful amplification. Optimized polymerase formulations can enable robust Extraction-Free PCR experiments. For example, Hemo KlenTaq can tolerate up to 20% whole blood added directly to a PCR mixture.

    Next Generation Sequencing Library Construction: Minimizing bias is important for even amplification of NGS libraries including GC-rich and AT-rich amplicons. NEBNext® High-Fidelity 2X PCR Master Mix (M0541) combines high fidelity and robust, even amplification making it an ideal choice for your NGS library construction.

    Multiplex PCR: Multiplex PCR involves the simultaneous amplification of two or more primer sets in a single reaction. Specialized enzyme formulations can also increase multiplex performance and reduce the need for troubleshooting. NEB offers both Taq-based and Q5-based formulations that are appropriate for use in multiplex PCR.

    Specialty PCR includes these areas of focus:

    Hot Start PCR
    Long Range PCR
    Extraction-Free PCR
    Multiplex PCR
    Bisulfite Sequencing
    Polymerases for NGS Library Preparation
    Fast PCR

    Protocols for Specialty PCR