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 (~280X Taq).
Bisulfite-converted DNA used in epigenetic analysis contains uracil. Use of polymerases like EpiMark® Hot Start Taq (NEB #M0490) that have been optimized to amplify bisulfite-converted and AT-rich amplicons ensures robust amplification.
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 (NEB #M0332) 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 (NEB #M0541) combines high fidelity and robust, even amplification making it an ideal choice for your NGS library construction.
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.
- Protocol for OneTaq 2X Master Mix with GC Buffer (M0483)
- Protocol for OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer (M0488)
- OneTaq® Quick-Load® 2X Master Mix with GC Buffer (M0487)
- PCR Protocol Phusion® DNA Polymerase
- Protocol for a Routine Taq PCR
- Taq DNA Polymerase Guidelines for PCR Optimization
- PCR Protocol for Taq DNA Polymerase
- PCR Protocol for Taq DNA Polymerase with Standard Taq (Mg-free) Buffer (M0320)
- PCR Protocol for Taq DNA Polymerase with ThermoPol Buffer (M0267)
- PCR Protocol for LongAmp® Hot Start Taq DNA Polymerase (M0534)
- Protocol for LongAmp™ Taq PCR Kit
- Protocol for Phusion® Hot Start Flex 2X Master Mix
- PCR Using NEBNext® High-Fidelity 2X PCR Master Mix (M0541)
- PCR Using Q5® Hot Start High-Fidelity DNA Polymerase (M0493)
- Protocol for Q5® Hot Start High-Fidelity 2X Master Mix
- Protocol for OneTaq Hot Start DNA Polymerase (M0481)
- Protocol for LongAmp™ Taq 2X Master Mix
- PCR Protocol for Hemo KlenTaq®
- Protocol for OneTaq® 2X Master Mix with Standard Buffer (M0482)
- Protocol for OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer
- PCR Protocol for Phusion® Hot Start Flex DNA Polymerase (M0535)
- Protocol Phusion® High-Fidelity PCR Master Mix with HF Buffer
- Protocol for OneTaq Hot Start 2X Master Mix with GC Buffer (M0485)
- Protocol for OneTaq Hot Start 2X Master Mix with Standard Buffer (M0484)
- PCR Optimization with Phusion® High-Fidelity PCR Kit
- Protocol for a Routine PCR with Phusion® High-Fidelity PCR Kit
- Guidelines for PCR Optimization with OneTaq® and OneTaq® Hot Start DNA Polymerases
- Protocol for OneTaq® Quick-Load 2X Master Mix with Standard Buffer (M0486)
- PCR Optimization of the Control Template using Phusion® High-Fidelity PCR Kit
- First strand cDNA synthesis OneTaq® RT-PCR Kit
- PCR Amplification with OneTaq® RT-PCR Kit
- Multiplex PCR Guidelines for Multiplex PCR 5X Master Mix
- Protocol for Phusion® High-Fidelity PCR Master Mix with GC Buffer
- Protocol for Quick-Load® Taq 2X Master Mix
- Protocol for Taq 2X Master Mix (M0270)
- Protocol for Taq 5X Master Mix (M0285)
- Quick Protocol for Multiplex PCR 5X Master Mix
- EpiMark® Hot Start Taq DNA Polymerase Guidelines for PCR (M0490)
- Protocol for a PCR reaction using Hot Start Taq 2X Master Mix (M0496)
- PCR Using Hot Start Taq DNA Polymerase (M0495)
- PCR Protocol for OneTaq® DNA Polymerase (M0480)
- PCR Protocol for LongAmp® Taq DNA Polymerase (M0323)
- Protocol for LongAmp™ Hot Start Taq 2X Master Mix
- Protocol for Q5® High-Fidelity 2X Master Mix
- PCR Optimization (E0555)
- Protocol for a Routine PCR (E0555)
- A-Tailing with Taq Polymerase
- PCR Using Q5® High-Fidelity DNA Polymerase (M0491)
- Loop-mediated Isothermal Amplification (LAMP)
- Protocol for a PCR reaction using NEBNext® Q5® Hot Start HiFi PCR Master Mix (M0543)
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High-fidelity, multiplex, and Fast PCR are just a few of the types of amplification reactions you may need to conduct. NEB offers a variety of DNA polymerases for all of your experimental needs. Breton explains which polymerases to choose.