DNA Amplification, PCR & qPCR
As the Polymerase Chain Reaction (PCR) is the most common DNA amplification method in molecular biology, NEB’s product portfolio features a large selection of polymerases geared towards this powerful method. As the first company to sell Taq DNA Polymerase to the research market, the first to discover a PCR-stable, high-fidelity DNA polymerase, and the first to provide reagents for PCR performed in space, NEB has a long history of developing reliable and convenient PCR tools. This commitment has continued with the recent development of OneTaq® DNA Polymerase for robust routine PCR and Q5® High-Fidelity DNA Polymerase for robust, ultra high-fidelity PCR (>280 X Taq 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, Taq and Q5, also benefit from novel aptamer-based hot start technology that does not require a separate activation step.
Despite the ubiquitous nature of PCR and qPCR, 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, to separate duplex DNA. To address some of the limitations of current isothermal amplification techniques, NEB has developed the next generation Bst, Bst 2.0 and a WarmStart® version of this enhanced polymerase, which enables room temperature reaction setup, yet is fully active at temperatures greater than 50°C.
RNA molecules can also be detected and manipulated through amplification via the use of reverse transcriptases (RT), which are RNA-dependent DNA Polymerases. RTs polymerize a strand of DNA that is complimentary to the original RNA template and is referred to as cDNA. This cDNA can then be further amplified through PCR, qPCR or isothermal methods as outlined above or detected in a single reaction using one-step RT-qPCR or RT-LAMP.
Nucleic acid amplification is a foundational process in molecular biology and, as a testament to its utility, new protocols and modifications are being developed constantly. At NEB, our goal is to use our understanding of enzymology, and dedication to providing high-quality products, to offer reagents for all of your applications. Please visit the application pages below to learn more.
TaqMan® is a registered trademark of Roche Molecular Systems, Inc.
SYBR® is a registered trademark of Molecular Probes, Inc.
- Isothermal Amplification
- Loop-Mediated Isothermal Amplification
- Whole Genome Amplification & Multiple Displacement Amplification
- Strand Displacement Amplification & Nicking Enzyme Amplification Reaction
- Helicase-dependent Amplification
- Recombinase Polymerase Amplification and SIBA
- Nucleic Acid Sequenced Based Amplification and Transcription Mediated Amplification
- Routine PCR
- High-Fidelity PCR
- PCR & Reaction Cleanup
- Polymerases for DNA Manipulation
- qPCR & RT-qPCR
- Dye-based qPCR & RT-qPCR
- Probe-based qPCR & RT-qPCR
- RT-PCR & cDNA Synthesis
- cDNA Synthesis
- Site Directed Mutagenesis
- Specialty PCR
- Extraction-Free PCR
- Hot Start PCR
- Long Range PCR
- Fast PCR
- Multiplex PCR
- Bisulfite Sequencing
- Polymerases for NGS Library Preparation
- Whole Genome Amplification
- 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 Q5® High-Fidelity 2X Master Mix
- PCR Optimization (E0555)
- Protocol for a Routine PCR (E0555)
- PCR Using Q5® High-Fidelity DNA Polymerase (M0491)
- Protocol for a PCR reaction using NEBNext® Q5® Hot Start HiFi PCR Master Mix (M0543)
- Luna® Universal qPCR Master Mix Protocol (#M3003)
- Luna® Universal One-Step RT-qPCR Kit Protocol (E3005)
- Luna® Universal Probe One-Step RT-qPCR Kit Protocol (E3006)
- Luna® Universal Probe qPCR Master Mix Protocol (M3004)
- Luna® Probe One-Step RT-qPCR Kit (No ROX) Protocol (NEB# E3007)
- Optimized Integration of New England Biolabs® Loop-mediated Isothermal Amplification (LAMP) Reagents with Axxin ISO Instruments
- Primer Monitor: an online tool to track SARS-CoV-2 variants that may impact primers used in diagnostic assays
- The Luna 4X RT-qPCR Mix and Luna SARS-CoV-2 RT-qPCR Multiplex Assay Kit enable high throughput, automated detection workflows
- Multiplex PCR using Q5 High-Fidelity DNA Polymerase
- Key considerations for optimal lyophilized reagent development
- Detection of the Omicron variant mutation at position 26,270 in the SARS-CoV-2 E gene using the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit
- Optimized conditions for the CDC Influenza SARS-CoV-2 (Flu SC2) Multiplex Assay using Luna One-Step RT-qPCR Reagents
- Multiplex real-time PCR detection of monkeypox virus using Luna® qPCR Reagents
- PCR Brochure
- DNA Polymerase Selection Chart
- Luna® One-Step RT-qPCR Troubleshooting Guide
- Luna® qPCR Troubleshooting Guide
- PCR Troubleshooting Guide
- Choosing RNA Input Amounts for NEB T2010
- General Guidelines for Successful RNA Purification Using the Monarch Total RNA Miniprep Kit
- Guidelines for PCR Optimization with OneTaq® and OneTaq® Hot Start DNA Polymerases
- Guidelines for PCR Optimization with Taq DNA Polymerase
- Guidelines for PCR Optimization with Thermophilic DNA Polymerases
- Optimization Tips for Luna® One-Step RT-qPCR
- Optimization Tips for Luna® qPCR
- Molecular Diagnostics for Gastrointestinal Parasites and Impact on Intestinal Microbiota in Rural Agentinian Children (2015)
- Amplification Reagents for Molecular Diagnostics Applications (2017)
- Colorimetric LAMP: Visual Detection for Simple Diagnostics (2017)
- A Single-tube, Low Input Protocol for Long Read Sequencing (2019)
- Gardner, A.F., Jackson, K.M., Boyle, M.M., Buss, J.A., Potapov, V., Gehring, A.M., Zatopek, K.M., Correa, I.R., Jr., Ong, J.L., Jack. W.E. (2019) Therminator DNA polymerase: Modified nucleotides and unnatural substrates Front Mol Biosci; 6, 28. PubMedID: 31069234, DOI: 10.3389/fmolb.2019.00028
- Poole, C.B., Sinha, A., Ettwiller, L., Apone, L., McKay, K., Panchapakesa, V., Lima, N.F., Ferreira, M.U., Wanji, S., Carlow, C.K.S (2019) In silico identification of novel biomarkers and development of new rapid diagnostic tests for the filarial parasites Mansonella perstans and Mansonella ozzardi Sci Rep; 9 (1), 10275. PubMedID: 31311985, DOI: 10.1038/s41598-019-46550-9
- Vladimir Potapov, Jennifer L. Ong. (2017) Examining Sources of Error in PCR by Single-Molecule Sequencing. PLOS One; PubMedID: 28683110
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Overview of PCR
This overview will walk you through how the Polymerase Chain Reaction (PCR) works.
Overview of qPCR
Learn the basics of qPCR in this short animation.
Types of PCR
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.
5 Tips for Setting Up Your PCR
Experiencing amplification frustration? Follow Melanie’s 5 quick and easy tips for PCR setup to improve your yields.
Why is Tm Important in Primer Design?
Why do primer melting temperatures matter? How is an annealing temperature calculated? Nicole has answers to these questions & more, in this quick video!