Hot Start PCR Reagents and Kits
Hot Start PCR Reagents and Kits
New England Biolabs, Inc. Phusion Hot Start Flex 2X Master Mix – 500 reactions (50 µl vol)
Phusion High-Fidelity DNA Polymerase offers high fidelity and robust performance, and thus can be used for all PCR applications. Its unique structure, a novel Pyrococcus-like enzyme fused with a processivity-enhancing domain, increases fidelity and speed. It is an ideal choice for cloning and can be used for long or difficult amplicons. With an error rate 50-fold lower than that of Taq DNA Polymerase and 6-fold lower than that of Pyrococcus furiosus DNA Polymerase, Phusion is one of the most accurate thermostable polymerases available. Phusion DNA Polymerase possesses 5' to 3' polymerase activity, 3' to 5' exonuclease activity and will generate blunt-ended products. Phusion Hot Start Flex 2X Master Mix offers robust high fidelity performance and room temperature reaction setup with the addition of an aptamer-based inhibitor. The formulation is supplied at a 2X concentration and contains dNTPs and Mg++, requiring only the addition of primers and DNA template for robust amplification.
New England Biolabs, Inc. OneTaq Hot Start DNA Polymerase – 200 units
OneTaq Hot Start DNA Polymerase is an optimized blend of Taq and Deep Vent DNA polymerases combined with an aptamer-based inhibitor. The 3' to 5' exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase and the hot start formulation combines convenience with decreased interference from primer dimers and secondary products. The OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields, regardless of a template's GC content. The inhibitor binds reversibly, blocking polymerase activity at temperatures below 45C. The polymerase is activated during normal cycling conditions, allowing reactions to be set up at room temperature. OneTaq Hot Start DNA Polymerase does not require a separate high temperature incubation step to activate the enzyme and can be used in typical Taq-based cycling protocols. It is supplied with two 5X buffers: (Standard and GC), as well as a High GC Enhancer solution.
New England Biolabs, Inc. WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) - 500 rxn
The WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) is designed to provide a simple one-step solution for Loop-Mediated Isothermal Amplification (LAMP) of DNA or RNA (RT-LAMP) targets. This kit includes the WarmStart Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) which contains a blend of Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase in an optimized buffer solution and a vial of LAMP Fluorescent Dye an intercalating dye provided at 50X concentration that can be used to track a LAMP reaction in real-time by fluorescence measurement. The inclusion of thermolabile UDG prevents carryover contamination where unintended product of a previous amplification can serve as the substrate of a subsequent reaction. Both Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase have been engineered for improved performance in LAMP and RT-LAMP reactions.
New England Biolabs, Inc. WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) - 100 rxn
The WarmStart Fluorescent LAMP/RT-LAMP Kit (with UDG) is designed to provide a simple one-step solution for Loop-Mediated Isothermal Amplification (LAMP) of DNA or RNA (RT-LAMP) targets. This kit includes the WarmStart Multi-Purpose LAMP/RT-LAMP 2X Master Mix (with UDG) which contains a blend of Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase in an optimized buffer solution and a vial of LAMP Fluorescent Dye an intercalating dye provided at 50X concentration that can be used to track a LAMP reaction in real-time by fluorescence measurement. The inclusion of thermolabile UDG prevents carryover contamination where unintended product of a previous amplification can serve as the substrate of a subsequent reaction. Both Bst 2.0 WarmStart DNA Polymerase and WarmStart RTx Reverse Transcriptase have been engineered for improved performance in LAMP and RT-LAMP reactions.
New England Biolabs, Inc. NEBNext Q5 Hot Start HiFi PCR Master Mix – 250 reactions
NEBNext Q5 Hot Start HiFi PCR Master Mix is the original hot start NEBNext formulation of Q5 High-Fidelity DNA polymerase. This formulation is also included as a component in all of the original NEBNext Ultra kits, for DNA and RNA library preparation. However, the NEBNext Ultra II Q5 Master Mix is now available, and this more recent hot start formulation further improves the uniformity of amplification of libraries, including superior performance with GC-rich regions.
New England Biolabs, Inc. NEBNext Q5U™ Master Mix – 250 reactions
NEBNext Q5U Master Mix is optimized for amplification of uracil-containing NGS libraries. This hot start formulation is designed to read through uracils, providing robust, high-fidelity and uniform library amplification. The polymerase component of the master mix, Q5U, is a modified version of Q5 High-Fidelity DNA Polymerase, a novel thermostable DNA polymerase that possesses 3´->5´ exonuclease activity, and is fused to a processivity-enhancing Sso7d domain. The NEBNext Q5U Master Mix is an aptamer-based hot start formulation that allows simple room temperature reaction set up. The convenient 2X master mix format contains dNTPs, Mg++ and a proprietary buffer, and requires only the addition of primers and DNA template for robust amplification.
New England Biolabs, Inc. NEBNext Q5U™ Master Mix – 50 reactions
NEBNext Q5U Master Mix is optimized for amplification of uracil-containing NGS libraries. This hot start formulation is designed to read through uracils, providing robust, high-fidelity and uniform library amplification. The polymerase component of the master mix, Q5U, is a modified version of Q5 High-Fidelity DNA Polymerase, a novel thermostable DNA polymerase that possesses 3´->5´ exonuclease activity, and is fused to a processivity-enhancing Sso7d domain. The NEBNext Q5U Master Mix is an aptamer-based hot start formulation that allows simple room temperature reaction set up. The convenient 2X master mix format contains dNTPs, Mg++ and a proprietary buffer, and requires only the addition of primers and DNA template for robust amplification.
New England Biolabs, Inc. Phusion Hot Start Flex DNA Polymerase – 100 units
Phusion High-Fidelity DNA Polymerase offers both high fidelity and robust performance, and thus can be used for all PCR applications. Its unique structure, a novel Pyrococcus-like enzyme fused with a processivity-enhancing domain, increases fidelity and speed. Phusion DNA Polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. With an error rate 50-fold lower than that of Taq DNA Polymerase and 6-fold lower than that of Pyrococcus furiosus DNA Polymerase, Phusion is one of the most accurate thermostable polymerases available. Phusion DNA Polymerase possesses 5' to 3' polymerase activity, 3' to 5' exonuclease activity and will generate blunt-ended products. The addition of an aptamer-based inhibitor allows room temperature reaction setup. Phusion Hot Start Flex DNA Polymerase is supplied with standard 5X Phusion HF Buffer, as well as 5X Phusion GC Buffer, which can be used for complex or GC-rich templates.
New England Biolabs, Inc. NheI-HF – 5000 units
A high fidelity restriction endonuclease that recognizes the sequence G^CTAGC. High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes.
New England Biolabs, Inc. OneTaq Hot Start 2X Master Mix with Standard Buffer – 500 reactions (50 µl vol)
OneTaq Hot Start 2X Master Mix with Standard Buffer is an optimized blend of Taq and Deep Vent DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to routine PCR applications from a variety of templates, including pure DNA solutions, bacterial colonies and cDNA products. The 3' to 5' exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase. The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient master mix formulation contains dNTPs, MgCl2 and other buffer components and stabilizers listed below, requiring only the addition of primers and DNA template for robust amplification.
New England Biolabs, Inc. OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer – 500 reactions (50 µl vol)
OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer is an optimized, ready-to-use blend of Taq and Deep Vent DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to routine PCR applications from a variety of templates, including pure DNA solutions, bacterial colonies and cDNA products. The 3' to 5' exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase. The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient Quick-Load master mix formulation contains dNTPs, MgCl2, buffer components and stabilizers as well as two commonly used tracking dyes for DNA gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~4 kb and Tartrazine migrates at ~10 bp. Both dyes are present in concentrations that do not mask co-migrating DNA bands
New England Biolabs, Inc. OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer – 500 reactions (50 µl vol)
OneTaq Hot Start Quick-Load 2X Master Mix with GC Buffer is an optimized, ready-to-use blend of Taq and Deep Vent DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to PCR applications from GC-rich templates, including pure DNA solutions, bacterial colonies, and cDNA products. The 3' to 5' exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase. The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient master mix formulation contains dNTPs, MgSO4, buffer components and stabilizers as well as two commonly used tracking dyes for DNA gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~4 kb and Tartrazine migrates at ~10 bp. Both dyes are present in concentrations that do not mask co-migrating DNA bands.
New England Biolabs, Inc. LongAmp Hot Start Taq 2X Master Mix – 100 reactions (50 µl vol)
LongAmp Hot Start Taq 2X Master Mix contains a unique blend of aptamer-based Hot Start Taq and Deep Vent DNA Polymerases. The aptamer-based inhibitor binds reversibly to the enzyme, inhibiting polymerase activity at temperatures below 45C, but releases the enzyme during normal PCR cycling conditions. This permits assembly of PCR reactions at room temperature. The LongAmp Hot Start Taq 2X Master Mix does not require a separate high temperature incubation step to activate the enzyme. The 3' tp 5' exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Hot Start Taq. LongAmp Hot Start Taq DNA Polymerase offers two-fold higher fidelity than Hot Start Taq DNA Polymerase alone. The convenient master mix formulation is supplied at a 2X concentration and contains dNTPs and Mg++, requiring only the addition of primers and DNA template for robust amplification. A wide range of PCR products can be generated; up to 30 kb from lambda or human genomic DNA.
New England Biolabs, Inc. BtsCI – 2000 units
A restriction endonuclease that recognizes the sequence GGATG_NN^.
New England Biolabs, Inc. OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer – 100 reactions (50 µl vol)
OneTaq Hot Start Quick-Load 2X Master Mix with Standard Buffer is an optimized, ready-to-use blend of Taq and Deep Vent DNA Polymerases combined with an aptamer-based inhibitor. This enzyme blend is ideally suited to routine PCR applications from a variety of templates, including pure DNA solutions, bacterial colonies and cDNA products. The 3' to 5' exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase. The hot start nature of the enzyme offers convenience with decreased interference from primer dimers and secondary products. The convenient Quick-Load master mix formulation contains dNTPs, MgCl2, buffer components and stabilizers as well as two commonly used tracking dyes for DNA gels. On a 1% agarose gel in 1X TBE, Xylene Cyanol FF migrates at ~4 kb and Tartrazine migrates at ~10 bp. Both dyes are present in concentrations that do not mask co-migrating DNA bands
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