Shop All Hot Start PCR Reagents and Kits

AmpliTaq Gold™ DNA Polymerase with Gold Buffer and MgCl2 Applied Biosystems™

AmpliTaq Gold® DNA Polymerase is a chemically modified form of AmpliTaq® DNA Polymerase requiring thermal activation. Features of this enzyme:

• Automated chemical hot-start enzyme for increased specificity, sensitivity, and yield
• Time-released thermal activation improves sensitivity in low copy number amplifications
• Successful multiplex PCR saves time and reagents
• Includes GeneAmp® 10X PCR Gold Buffer with MgCl2

Hot-start activation
The modified enzyme is provided in an inactive state. Upon thermal activation, the modifier is permanently released, regenerating active enzyme. The resulting hot-start PCR amplification provides increased sensitivity, specificity, and yield over conventional PCR techniques.

Higher specificity, higher yield
AmpliTaq Gold® DNA Polymerase can be activated partially or completely in a pre-PCR heat step, or can be allowed to activate slowly in a time-released manner during the denaturation steps of thermal cycling. With or without a limited up-front heat activation step, active enzyme is released slowly during thermal cycling to match template concentration and increase specificity. The yield of specific product increases because reactants are not wasted in the formation of unintended products. Because AmpliTaq Gold® DNA Polymerase is a chemical hot-start enzyme, there is no worry of biological contamination.

GeneAmp® 10X PCR Gold Buffer is formulated to provide flexible, efficient activation of AmpliTaq Gold® DNA Polymerase, resulting in a highly specific, robust PCR amplification. The ionic strength and the pH of GeneAmp® 10X PCR Gold Buffer have been optimized to provide a wider activation temperature and time range when used in conjunction with AmpliTaq Gold® DNA Polymerase.

AmpliTaq Gold® 360 DNA Polymerase for superior performance
Also available is AmpliTaq Gold® 360 DNA Polymerase for even better PCR performance. Compared to AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process to eliminate contaminating bacterial DNA sequences from the enzyme preparation. When used with the enhanced 360 Gold Buffer, this ultra-pure enzyme, in addition to its hot-start capabilities, reduces false positive results, amplifies low-level target sequences, and promotes the amplification of a variety of templates. Also included with this product is a 360 GC Enhancer to work through challenging GC-rich templates.

Phusion Green Hot Start II High-Fidelity PCR Master Mix Thermo Scientific™

Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR. Featuring an error rate 50-fold lower than that of Taq, and 6-fold lower than that of Pfu, Phusion High-Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity. Phusion DNA Polymerases offer robust performance with short protocol times, even in the presence of PCR inhibitors, and generate higher yields with lower enzyme amounts than other DNA polymerase.

Highlights

• Reaction set up at room temperature
• No non-specific amplification and primer degradation during reaction set up
• Zero-time reactivation due to unique hot start technology
• High fidelity (52X Taq)
• Fast PCR due to short extension times (15-30 s/kb)
• Robust reactions, minimal optimization needed
• Increased product yields with minimal enzyme amounts
• Direct loading on gels

With Phusion Hot Start II High-Fidelity DNA Polymerases amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand that inhibits DNA polymerase activity at room temperature. The Affibody ligand also inhibits the 3´→5´ exonuclease activity of the polymerase, thus preventing degradation of primers and template DNA during reaction set up. At temperatures that promote polymerase activity, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols.
Phusion Green Hot Start II High-Fidelity PCR Master Mix is convenient 2X mix designed to minimize the number of pipetting steps. The master mix contains Phusion Hot Start II DNA Polymerase, nucleotides and optimized reaction buffer including MgCl2. The buffer also includes a density reagent and two tracking dyes for direct loading of PCR products on a gel.

Applications

• High-fidelity PCR
• High throughput
• Amplification of difficult (GC-rich) templates
• Template generation for sequencing
• Multiplex PCR
• Long-range PCR
• Cloning
• Mutagenesis
• Microarray

Using Phusion DNA Polymerases
Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases). For optimal results, use our Tm calculator at www.thermofisher.com/tmcalculator.

Phire Hot Start II DNA Polymerase Thermo Scientific™

Thermo Scientific Phire Hot Start II DNA Polymerase is an enhanced PCR enzyme for routine and high throughput PCR applications. It outperforms every Taq-based hot start polymerase on the market. Phire Hot Start II DNA Polymerase is significantly faster, extremely robust, and also capable of amplifying long DNA fragments with high yields. These features are achieved through advanced protein engineering of the polymerase. It incorporates a unique double-stranded DNA binding domain which allows short extension times (10 to 15 s/kb), improves yields, and increases fidelity 2-fold compared to Taq DNA polymerase.

The hot start modification of Phire Hot Start II DNA Polymerase is based on the Affibody inactivation technology. This technology increases the specificity of the PCR reaction with no additional time required for initial activation of the enzyme.

Highlights

Quick hot start—No reactivation step
Fast enzyme—Amplify four times faster than with hot start Taq
Robust—Minimal reaction optimization due to high inhibitor tolerance
High yields—Abundant products due to high efficiency
Longer PCR products—Amplify significantly longer DNA fragments than with any hot start Taq

Applications

• Hot Start PCR
• Routine PCR
• Non-high fidelity PCR
• Fast PCR
• High throughput PCR
• Genotyping

Note:The optimal annealing temperature for Phire DNA Polymerases may differ significantly from that of Taq-based polymerases. For optimal results start by accurately calculating your Tm with our Tm calculator.

Phire Green Hot Start II DNA Polymerase is also available (F-124S or F-124L) for direct loading of PCR products on gel.

Platinum™ SuperFi II Green PCR Master Mix Invitrogen™

Invitrogen Platinum SuperFi II Green PCR Master Mix (2X) contains a ready-to-use mixture of Platinum SuperFi II DNA Polymerase, Platinum SuperFi II Buffer, and dNTPs for convenient PCR setup, as well as two tracking dyes for direct loading of PCR products on gels. Platinum SuperFi II DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with innovative SuperFi II Buffer, enabling universal primer annealing for the highest success in PCR. It is ideally suited for cloning, mutagenesis, and other applications benefiting from supreme sequence accuracy.

Features of Platinum SuperFi II Green PCR Master Mix include:
• Exceptional >300X Taq fidelity
• Universal primer annealing at 60°C
• Superior specificity, sensitivity, and yields
• Robust amplification of difficult-to-amplify targets (e.g., those with suboptimal purity, ˃65% GC content, long PCR requirement)

Platinum SuperFi II DNA Polymerase is an engineered enzyme with high processivity and increased resistance to PCR inhibitors. It also enables fast-cycling protocols and amplification of long targets (up to 20 kb). Platinum hot-start technology is based on proprietary antibodies that inhibit enzyme activity until the initial PCR denaturation step, preventing non-specific amplification and primer degradation. This technology also enables reaction set up at room temperature and provides increased sensitivity and yield.

Due to the unique composition of the SuperFi II PCR buffer, the annealing temperature is 60°C for most primer pairs designed following the general design rules. Isostabilizing molecules in the buffer increase primer-template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize the annealing temperature for each primer pair. With Platinum SuperFi II DNA Polymerase, different PCR assays can be cycled together using the same protocol with a universal primer annealing temperature and an extension step selected for the longest fragment to be amplified.

Applications of Platinum SuperFi II DNA Polymerase:
• High-fidelity PCR
• Cloning and sub-cloning
• Site-directed mutagenesis
• Amplification of GC-rich templates
• Template generation for sequencing
• High-throughput PCR
• Amplification of samples with suboptimal purity
• Long PCR
• Fast PCR

Platinum SuperFi II PCR Master Mix is also available, which is the same master mix without the tracking dyes.

View more information about Platinum SuperFi II DNA Polymerase products ›

Platinum™ Green Hot Start PCR Master Mix (2X) Invitrogen™

Invitrogen Platinum Green Hot Start PCR Master Mix offers Platinum Taq DNA polymerase in an optimized PCR buffer with magnesium and dNTPs for convenient PCR setup. It retains all the features of Platinum Taq DNA Polymerase: specificity, robustness, and reliability. Platinum Green Hot Start PCR Master Mix is supplemented with two tracking dyes for the convenience of direct gel loading of PCR products.

Product features:
• High specificity and increased yields with antibody-mediated hot-start PCR
• Convenient room-temperature reaction setup
• Direct gel loading with green PCR master mix
• Versatile formulation for broad range of amplicons

About Platinum Taq DNA Polymerase
Platinum Taq DNA Polymerase is a recombinant Taq DNA polymerase complexed with proprietary antibodies that inhibit enzyme activity at room temperature, reducing nonspecific target amplification at lower temperatures. The antibodies dissociate during the initial PCR denaturation step and DNA polymerase regains its full activity. Just as with Taq DNA Polymerase, Platinum Taq DNA Polymerase has a non-template-dependent terminal transferase activity that adds a 3' deoxyadenosine to product ends and has a 5'→3' exonuclease activity.

Using Platinum Green Hot Start PCR Master Mix
Convenient Platinum Green Hot Start PCR Master Mix provides a ready-to-use mixture of DNA polymerase, salts, magnesium, and dNTPs, thus reducing the number of pipetting steps during PCR reaction setup. The two tracking dyes in Platinum Green Hot Start PCR Master Mix enable direct gel loading of PCR products, thus eliminating tedious steps of dye addition. Platinum Green Hot Start PCR Master Mix is supplied with a separate vial of Platinum GC Enhancer formulated for specific amplification and improved yields of GC-rich targets. Use Platinum Green Hot Start PCR Master Mix in PCR applications such as genotyping, gene expression profiling, and next-generation sequencing.

Note: For superior PCR performance, we recommend next generation enzyme Platinum II Taq Hot-start DNA Polymerase. Platinum II Taq Hot-Start DNA Polymerase is designed for universal primer annealing and fast, easy PCR with its unique combination of innovative buffer, high-performance engineered Taq DNA polymerase, and superior hot-start technology. Platinum II Hot-Start Green PCR Master Mix contains Platinum II Taq Hot-Start DNA Polymerase in a ready-to-use mixture with Platinum II PCR buffer and dNTPs, thus reducing the number of pipetting steps during PCR reaction setup. It additionally contains a density reagent and two tracking dyes for direct loading of PCR products on gels.

Phire Green Hot Start II PCR Master Mix Thermo Scientific™

Thermo Scientific Phire Green Hot Start II PCR Master Mix is convenient 2X mix designed to minimize the number of pipetting steps. The master mix contains Phire Hot Start II DNA Polymerase, nucleotides and optimized reaction buffer including MgCl2. Only template and primers need to be added to PCR reaction. The master mix also includes a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with enzyme performance and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.

Phire Hot Start II DNA Polymerase, inluded in the master mix, is an enhanced PCR enzyme for routine and high throughput PCR applications. It is significantly faster, extremely robust, and also capable of amplifying long DNA fragments with high yields. These features are achieved through advanced protein engineering of the polymerase. It incorporates a unique double-stranded DNA binding domain which allows short extension times (10 to 15 s/kb), improves yields, and increases fidelity 2-fold compared to Taq DNA polymerase.

Highlights

Quick hot start—No reactivation step
Fast enzyme—Amplify four times faster than with hot start Taq
Robust—Minimal reaction optimization due to high inhibitor tolerance
High yields—Abundant products due to high efficiency
Longer PCR products—Amplify significantly longer DNA fragments than with any hot start Taq
Direct loading on gel

Applications

• Hot Start PCR
• Routine PCR
• Non-high fidelity PCR
• Fast PCR
• High throughput PCR
• Genotyping

Note:The optimal annealing temperature for Phire DNA Polymerases may differ significantly from that of Taq-based polymerases. For optimal results start by accurately calculating your Tm with our Tm calculator.

AmpliTaq Gold™ DNA Polymerase with Buffer I Applied Biosystems™

AmpliTaq Gold® DNA Polymerase is a chemically modified form of AmpliTaq® DNA Polymerase requiring thermal activation. Features of this enzyme:

• Automated chemical hot-start enzyme for increased specificity, sensitivity, and yield
• Time-released thermal activation improves sensitivity in low copy number amplifications
• Successful multiplex PCR saves time and reagents
• Includes GeneAmp® 10X PCR Buffer I containing 15 mM MgCl2

Hot-start activation
The modified enzyme is provided in an inactive state. Upon thermal activation, the modifier is permanently released, regenerating active enzyme. The resulting hot-start PCR amplification provides increased sensitivity, specificity, and yield over conventional PCR techniques.

Higher specificity, higher yield
AmpliTaq Gold® DNA Polymerase can be activated partially or completely in a pre-PCR heat step, or can be allowed to activate slowly in a time-released manner during the denaturation steps of thermal cycling. With or without a limited up-front heat activation step, active enzyme is released slowly during thermal cycling to match template concentration and increase specificity. The yield of specific product increases because reactants are not wasted in the formation of unintended products. Because AmpliTaq Gold® DNA Polymerase is a chemical hot-start enzyme, there is no worry of biological contamination.

AmpliTaq Gold® 360 DNA Polymerase for superior performance
Also available is AmpliTaq Gold® 360 DNA Polymerase for even better PCR performance. Compared to AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process to eliminate contaminating bacterial DNA sequences from the enzyme preparation. When used with the enhanced 360 Gold Buffer, this ultra-pure enzyme, in addition to its hot-start capabilities, reduces false positive results, amplifies low-level target sequences, and promotes the amplification of a variety of templates. Also included with this product is a 360 GC Enhancer to work through challenging GC-rich templates.

Platinum™ Direct PCR Universal Master Mix Invitrogen™

Invitrogen Platinum Direct PCR Universal Master Mix is designed to amplify DNA directly from various samples without the need to purify DNA. It contains high-performing, engineered Platinum II Taq Hot-Start DNA Polymerase with dNTPs in an innovative buffer that enables universal primer annealing for superior performance in direct PCR applications.

Features of Platinum Direct PCR Universal Master Mix include:
• Direct DNA amplification from various samples due to engineered, inhibitor-tolerant Platinum II Taq Hot-Start DNA Polymerase
• Universal primer annealing temperature due to innovative buffer that enables primer annealing at 60°C
• Superior specificity, sensitivity, and yields due to Platinum hot-start technology

Platinum Direct PCR Universal Master Mix is designed to work with a variety of samples of different origins such as animal, human, plant, insect, worm, and bacterial samples. The kit includes optimized reagents to achieve superior results. The quick lysis protocol enables efficient amplification from templates up to 8 kb in size and co-cycling of different length fragments. Samples in the lysis buffer can be stored for later use. Platinum Direct PCR Universal Master Mix is provided with the optional Platinum GC Enhancer for specific amplification and improved yields of GC-rich targets.

Engineered Platinum II Taq DNA polymerase confers faster cycling and tolerance to reaction inhibitors originating from sample material. Proprietary Platinum Taq antibodies block polymerase activity at ambient temperatures and dissociate after the initial denaturation step. This automatic 'hot start' provides increased sensitivity, specificity, and yield, while allowing reaction assembly at room temperature.

Due to the unique composition of the Platinum Direct PCR Universal Master Mix, the annealing temperature of 60°C can be used for most primer pairs that follow the general design rules. Isostabilizing molecules in the buffer increase primer-template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize annealing temperature for each primer pair. Different PCR assays can be cycled together using the same protocol with a universal primer annealing temperature and an extension step selected for the longest fragment to be amplified.

View more information about Platinum Direct PCR Universal Master Mix ›

GeneAmp™ 10X PCR Gold Buffer & MgCl2 Applied Biosystems™

Applied Biosystems GeneAmp 10X PCR Gold Buffer is formulated to provide flexible, efficient activation of AmpliTaq Gold DNA Polymerase, resulting in highly specific, robust PCR amplification.

The ionic strength and pH of GeneAmp 10X PCR Gold Buffer have been optimized to provide a wider activation temperature and time range when used in conjunction with AmpliTaq Gold DNA Polymerase.

GeneAmp 10X PCR Gold Buffer is both magnesium ion-free and gelatin-free. A separate MgCl2 solution is provided. When used together, GeneAmp 10X PCR Gold Buffer and the MgCl2 solution offer more versatility in optimizing the magnesium ion concentration to achieve optimal PCR amplification with any specific set of primers and DNA template.

Platinum™ Taq DNA Polymerase Invitrogen™

Invitrogen Platinum Taq DNA Polymerase is a convenient and reliable 'hot start' thermostable DNA polymerase for PCR that provides enhanced specificity over that of Taq DNA Polymerase. The 'hot start' property of the enzyme is conferred by thermolabile monoclonal antibodies that render Taq DNA polymerase inactive until the initial PCR denaturation step, thus preventing the extention of nonspecifically annealed primers and improving product yield.

Using Platinum Taq DNA Polymerase
The hot-start property of Platinum Taq DNA Polymerase allows for convenient reaction assembly at room temperature. Just as with Taq DNA Polymerase, Platinum Taq DNA Polymerase has a non-template–dependent terminal transferase activity that adds a 3′ deoxyadenosine to product ends, and has a 5′→3′ exonuclease activity. PCR products generated with Platinum Taq DNA Polymerase may be used in the same downstream applications without protocol modifications. Use Platinum Taq DNA Polymerase for the amplification of DNA from complex genomic, viral, and plasmid templates, as well as in RT-PCR.

Note: For superior PCR performance, we recommend next-generation enzyme Platinum II Taq Hot-start DNA Polymerase. Platinum II Taq Hot-Start DNA Polymerase is designed for universal primer annealing and fast, easy PCR with its unique combination of innovative buffer, high-performance engineered Taq DNA polymerase, and superior hot-start technology.

Platinum™ II Taq Hot-Start DNA Polymerase Invitrogen™

Invitrogen Platinum II Taq Hot-Start DNA Polymerase is designed for universal primer annealing and fast, easy PCR with its unique combination of innovative buffer, high-performance engineered Taq DNA polymerase, and superior hot-start technology.

Features of Platinum II Taq Hot-Start DNA Polymerase include:
Innovative buffer—enables universal annealing temperature by isostabilizing primer-template duplex structures
Engineered Taq DNA polymerase—confers fast cycling and resistance to common inhibitors
Platinum hot-start technology—enables superior specificity, sensitivity, and yields; allows for room temperature reaction setup

Platinum II Taq Hot-Start DNA Polymerase is an engineered Taq DNA polymerase that shows increased resistance to reaction inhibitors originating from sample material or DNA purification steps. The polymerase has a higher DNA synthesis rate and delivers PCR results more than two times faster than other Taq DNA polymerases. Proprietary Platinum Taq antibodies block polymerase activity at ambient temperatures and dissociate after the initial denaturation step at 94°C. This automatic 'hot start' provides increased sensitivity, specificity, and yield, while allowing reaction assembly at room temperature.

Due to the unique composition of the Platinum II PCR buffer, the annealing temperature is 60°C for most primer pairs designed following the general design rules. Isostabilizing molecules in the buffer increase primer–template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize annealing temperature for each primer pair. With Platinum II Taq Hot-Start DNA Polymerase, different PCR assays can be cycled together using the same protocol with universal primer annealing temperature and the extension step selected for the longest fragment to be amplified.

Platinum II Taq Hot-Start DNA Polymerase is provided with the optional Platinum GC Enhancer for specific amplification and improved yields of GC-rich targets.

Use Platinum II Taq Hot-Start DNA Polymerase for the amplification of DNA from complex genomic, viral, and plasmid templates, as well as in RT-PCR, in applications like genotyping, high-throughput PCR, or with samples of suboptimal purity.

For increased convenience, we offer Platinum II Hot-Start PCR Master Mix (2X), where Platinum II Taq Hot-Start DNA Polymerase is provided in a ready-to-use mixture with Platinum II PCR buffer and dNTPs, thus reducing the number of pipetting steps during PCR reaction setup. Platinum II Hot-Start Green PCR Master Mix (2X) is also available, which additionally contains a density reagent and two tracking dyes for direct loading of PCR products on gels, further streamlining the PCR workflow from setup to final analysis of the result.

Find out more at www.thermofisher.com/platinumiitaq ›

PCR Master Mix Starter Pack Invitrogen™

The PCR Master Mix Starter Pack is composed of three different PCR master mixes for easy evaluation of their suitability for use in your research. These master mixes provide our best DNA polymerases for routine PCR, hot-start PCR, and high-fidelity PCR needs. Thermo Scientific DreamTaq Green PCR Master Mix contains an enhanced Taq DNA polymerase optimized for all standard PCR applications. Invitrogen Platinum II Hot Start PCR Master Mix contains hot-start Taq DNA polymerase for specific, robust, and reliable PCR. Invitrogen SuperFi II PCR Master Mix features superior performance and an error rate 300X lower than that of Taq. It is an excellent choice for cloning, mutagenesis, and other applications requiring high fidelity.

Master mixes included in this starter pack:
K1081 DreamTaq Green PCR Master Mix (2X), 200 reactions
14000012 Platinum II Hot Start PCR Master Mix (2X), 50 reactions
12368010 Platinum SuperFi II PCR Master Mix, 100 reactions

Platinum™ Taq DNA Polymerase, DNA-free Invitrogen™

Invitrogen Platinum Taq DNA Polymerase, DNA-free, is manufactured using a novel, closed single-use system together with stringent quality-control testing to minimize the risk of contamination with bacterial and human DNA. This enzyme is an ideal choice for PCR-based applications requiring the highest sensitivity without false-positive results from reagent-borne contamination. This DNA-free enzyme provides the same high level of performance and lot-to-lot consistency as standard Platinum Taq DNA Polymerase.

DNA-free Platinum Taq DNA Polymerase offers:
• Certified low-DNA contamination level (≤0.01 copy of bacterial gDNA per enzyme unit)
• Confidence in qualitative and quantitative detection of low-abundance microorganisms due to signal absence with no-template/reagents-only PCR controls
• Same performance as Platinum Taq DNA Polymerase manufactured using conventional process

Platinum Taq DNA Polymerase, DNA-free, is a recombinant Taq polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures and dissociates after the initial denaturation step at 94°C. This automatic “hot start” provides increased sensitivity, specificity, and yield, while allowing convenient reaction assembly at room temperature. Just as with Taq DNA Polymerase, Platinum Taq DNA Polymerase, DNA-free, has a non-template–dependent terminal transferase activity that adds a 3′ deoxyadenosine to product ends, and has a 5′→3′ exonuclease activity.

Use Platinum Taq DNA Polymerase, DNA-free, for the amplification of DNA from complex genomic and viral templates, as well as in RT-PCR. It is the perfect choice for PCR assays where higher sensitivity and specificity are needed to minimize ambiguous or false-positive results. Find out more at www.thermofisher.com/dna-free.

AmpliTaq Gold™ DNA Polymerase with Buffer II and MgCl2 Applied Biosystems™

AmpliTaq Gold® DNA Polymerase is a chemically modified form of AmpliTaq® DNA Polymerase requiring thermal activation. Features of this enzyme:

• Automated chemical hot-start enzyme for increased specificity, sensitivity, and yield
• Time-released thermal activation improves sensitivity in low copy number amplifications
• Successful multiplex PCR saves time and reagents
• Includes GeneAmp® 10X PCR Buffer II with MgCl2

Hot-start activation
The modified enzyme is provided in an inactive state. Upon thermal activation, the modifier is permanently released, regenerating active enzyme. The resulting hot-start PCR amplification provides increased sensitivity, specificity, and yield over conventional PCR techniques.

Higher specificity, higher yield
AmpliTaq Gold® DNA Polymerase can be activated partially or completely in a pre-PCR heat step, or can be allowed to activate slowly in a time-released manner during the denaturation steps of thermal cycling. With or without a limited up-front heat activation step, active enzyme is released slowly during thermal cycling to match template concentration and increase specificity. The yield of specific product increases because reactants are not wasted in the formation of unintended products. Because AmpliTaq Gold® DNA Polymerase is a chemical hot-start enzyme, there is no worry of biological contamination.

AmpliTaq Gold® 360 DNA Polymerase for superior performance
Also available is AmpliTaq Gold® 360 DNA Polymerase for even better PCR performance. Compared to AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process to eliminate contaminating bacterial DNA sequences from the enzyme preparation. When used with the enhanced 360 Gold Buffer, this ultra-pure enzyme, in addition to its hot-start capabilities, reduces false positive results, amplifies low-level target sequences, and promotes the amplification of a variety of templates. Also included with this product is a 360 GC Enhancer to work through challenging GC-rich templates.

Phusion Hot Start II High-Fidelity PCR Master Mix Thermo Scientific™

Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR. Featuring an error rate 50-fold lower than that of Taq, and 6-fold lower than that of Pfu, Phusion High-Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity. Phusion DNA Polymerases offer robust performance with short protocol times, even in the presence of PCR inhibitors, and generate higher yields with lower enzyme amounts than other DNA polymerase.

Highlights

• Reaction set up at room temperature
• No non-specific amplification and primer degradation during reaction set up
• Zero-time reactivation due to unique hot start technology
• High fidelity (52X Taq)
• Fast PCR due to short extension times (15-30 s/kb)
• Robust reactions, minimal optimization needed
• Increased product yields with minimal enzyme amounts
• Available as a Green buffer format permitting direct loading on gels (F-566S or F-566L)

Using Phusion Hot Start II High-Fidelity DNA Polymerase, amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand that inhibits DNA polymerase activity at room temperature. The Affibody ligand also inhibits the 3´→5´ exonuclease activity of the polymerase, thus preventing degradation of primers and template DNA during reaction set up. At temperatures that promote polymerase activity, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols. Phusion Hot Start II High-Fidelity PCR Master Mix is convenient 2X mix designed to minimize the number of pipetting steps. The master mix contains Phusion Hot Start II DNA Polymerase, nucleotides and optimized reaction buffer including MgCl2. Only template and primers need to be added to the PCR reaction.

Applications

• High-fidelity PCR
• High throughput
• Amplification of difficult (GC-rich) templates
• Template generation for sequencing
• Multiplex PCR
• Long-range PCR
• Cloning
• Mutagenesis
• Microarray

Using Phusion DNA Polymerases
Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases). For optimal results, use our Tm calculator at www.thermofisher.com/tmcalculator.

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