Shop All PCR & Cloning Enzymes

Phire Green 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 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.

Phire Green format is a combination of Phire Hot Start II DNA Polymerase and 5X Phire Green Buffer. The buffer 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.

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 with Green Buffer

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 Hot Start II PCR Master Mix (Thermo Scientific™)

Thermo Scientific Phire 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.

Phire Hot Start II DNA Polymerase, inluded in the master mix, 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.

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.

Anza™ 23 PstI (Invitrogen™)

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3'  G ↑A  C  G  T  C   5' 

Invitrogen™ Anza™ 23 PstI is a restriction enzyme that cuts DNA at this recognition site: CTGCA^G, completely digesting the DNA in 15 minutes at 37°C.

For superior convenience, a single buffer and protocol are used with all Anza™ restriction enzymes. The single Anza buffer allows multiple restriction enzymes to be used simultaneously in one reaction. This helps eliminate the need for sequential digests and the DNA clean-up necessary when changing buffers. Additionally, the Anza buffer and restriction enzymes have been designed for flexibility in digestion times. Digestions of up to 16 hours can be carried out without star activity.

Convenient—all Anza enzymes are 100% functional in the single Anza buffer
Fast—complete digestion in 15 minutes
Flexible—overnight digestion without star activity
Simple—a single digestion protocol for all DNA types
Complete system—incorporates Anza DNA Modifying Enzymes for a complete cloning experience

Anza DNA Modifying Enzymes include the Anza T4 DNA Ligase Master Mix, Anza Alkaline Phosphatase Kit, Anza T4 PNK Kit, Anza DNA Blunt End Kit, and Anza DNA End Repair Kit. Use these enzymes for a complete cloning workflow.

Anza Red Buffer, which contains a density gradient and tracking dyes, is supplied for added convenience. Digestion reactions are as efficient in the Anza Red Buffer as in the Anza Buffer. Use of the Anza Red Buffer helps reduce pipetting steps when preparing multiple digestion reactions and a subsequent step includes agarose gel electrophoresis. For applications that require product analysis by fluorescence excitation, the colorless Anza Buffer without the tracking dyes is recommended.

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.

Platinum™ GenoType Tsp DNA Polymerase (Invitrogen™)

Platinum® GenoType Tsp DNA Polymerase is designed and qualified specifically for amplification of dinucleotide repeat markers in PCR-based genotyping applications. The enzyme is pre-complexed with a thermolabile inhibitor containing monoclonal antibodies to Tsp DNA polymerase, providing an automatic "hot-start" method that improves PCR specificity and allows for room temperature reaction assembly. Platinum® GenoType Tsp DNA Polymerase:

• Exhibits minimal non-templated nucleotide addition to PCR products
• Lacks both 5´ and 3´ exonuclease activity
• Amplifies fragments up to 500 bp

Application
Amplification of dinucleotide repeat loci in applications in which elimination of non-templated nucleotide addition is desirable.

Unit definition
One unit of Platinum® GenoType Tsp DNA Polymerase has been functionally determined to be equivalent to one unit of Taq DNA Polymerase in amplification of dinucleotide repeats using standard Taq DNA polymerase reaction conditions. One Tsp DNA polymerase unit approximates 2.5 activity units. An activity unit incorporates 10 nmoles of deoxyribonucleotide into acid-precipitable material in 30 min at 74°C under optimized reaction conditions.

Eam1105I (AhdI) (10 U/µL) (Thermo Scientific™)

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Thermo Scientific Eam1105I (AhdI) restriction enzyme recognizes GACNNN^NNGTC sites and cuts best at 37°C in its own unique buffer (Isoschizomers: AhdI, AspEI, BmeRI, DriI, EclHKI). See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities

Applications

• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP

Note: Low salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.
For methylation sensitivity, refer to product specifications.

Phusion High-Fidelity PCR Kit (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

• High fidelity (52X Taq)
• Fast PCR due to short extension times (15-30 s/kb)
• Robust performance, minimal optimization needed
• High yields of PCR products with minimal enzyme amounts
• Available as a Green buffer format for direct loading of PCR products on gels (F-534S or F-534L)

Applications

• High-fidelity PCR
• Cloning
• Template generation for sequencing
• Amplification of difficult (GC-rich) templates
• Long-range PCR (up to 20 kb)
• Mutagenesis
• High throughput PCR
• Microarray
Thermo Scientific Phusion High-Fidelity PCR Kit contains all the necessary reagents for PCR including control lambda DNA template and primers for 1.3 kb and 10 kb amplicons. The DNA template amount is sufficient for performing 20 control reactions in 50 µL volume or 50 control reactions in 20 µL volume.

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.

FspBI (BfaI) (10 U/µL) (Thermo Scientific™)

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3'  G  A  T ↑C   5' 

Thermo Scientific FspBI (BfaI) restriction enzyme recognizes C^TAG sites and cuts best at 37°C in Tango buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion. Isoschizomers: BfaI, MaeI, XspI.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities

Applications

• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP

Note: FspBI cleavage is impaired by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain such as GM2163. For methylation sensitivity, refer to product specifications.

HhaI (10 U/µL) (Thermo Scientific™)

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3'  C ↑G  C  G   5' 

Thermo Scientific HhaI restriction enzyme recognizes GCG^C sites and cuts best at 37°C in Tango buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion. Isoschizomers: AspLEI, BstHHI, CfoI, HinP1I, HspAI.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities

Applications

• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP

Note: For methylation sensitivity, refer to product specifications.

AmpliTaq Gold™ 360 DNA Polymerase (Applied Biosystems™)

The Applied Biosystems® AmpliTaq Gold® 360 DNA Polymerase, when used with improved AmpliTaq Gold 360 Buffer, and the optional 360 GC Enhancer, amplifies a vast range of DNA sequence contexts. AmpliTaq Gold® 360 DNA Polymerase delivers 360° coverage for a full range of targets.

* Optimized for the broadest range of targets—from everyday to challenging
* Unmatched sensitivity, specificity, and yield
* Robust amplification of GC-rich sequences with market-leading 360 GC Enhancer
* Achieves the highest quality sequencing data

Optimized for Easy and Challenging Targets
Challenging targets include AT-rich, GC-rich, primer-dimer forming amplicons, homopolymer repeats, and amplicons that pose sequencing challenges. Amplicons that previously required specialized enzymes and reaction conditions can now be reproducibly amplified with a single reagent under standardized conditions (Figure 1). Competitive benchmarking across more than 40 amplicons distinguishes AmpliTaq Gold® 360 as the best-performing enzyme, ensuring the highest probability of success for the amplification of both everyday and challenging targets (Table 1).

As shown in Figure 1, GC-rich regions are especially poorly amplified with competitor DNA polymerases and the original AmpliTaq Gold, while AmpliTaq Gold 360 provides successful, robust amplification.

Optimized for Hot-Start PCR
AmpliTaq Gold 360 DNA Polymerase provides the same hot-start specificity as AmpliTaq Gold DNA polymerase. A high-temperature incubation step is required to activate AmpliTaq Gold DNA Polymerase, which ensures that the active enzyme is generated only at temperatures in which the DNA is fully denatured and when the primers are not annealed.
When the polymerase is added to the reaction mixture at room temperature, primer extension does not occur because the enzyme is inactivated. Any low stringency mispriming events that may have occurred will not be enzymatically extended and will not be amplified. Hence, PCR setup can be performed at room temperature without concern for extension at misprimed sites. The amount of AmpliTaq Gold 360 DNA polymerase increases in the reaction slowly with each cycle number, and specific product yield increases without buildup of nonspecific products, including primer dimers. Excellent specificity across a broad range of targets is shown in Figure 1 and is summarized in Figure 2. The exquisite specificity allows easier multiplexing and allelic discrimination.

The amount of AmpliTaq Gold 360 DNA polymerase increases in the reaction slowly with each cycle number, and specific product yield increases without buildup of nonspecific products, including primer dimers. Excellent specificity across a broad range of targets is shown in Figure 1 and is summarized in Figures 2A & B. The exquisite specificity allows easier multiplexing and allelic discrimination.

Superior Sensitivity and Amplification Length
Compared to the original AmpliTaq Gold® DNA Polymerase, AmpliTaq Gold® 360 DNA Polymerase is purified by an additional proprietary separation process which reduces 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 including those from bacterial and human genomes.

AmpliTaq Gold 360 DNA Polymerase efficiently amplifies targets present at low copy number (Figure 3), even in the presence of high concentrations of complex DNA, making it especially suited for low-copy pathogen detection, and amplification of targets from degraded DNA samples. The extreme purity of the enzyme contributes to its unmatched sensitivity. AmpliTaq Gold 360 DNA Polymerase efficiently and reproducibly amplifies long (up to 5 Kb) sequences. Figure 4 demonstrates robust PCR amplification of long human and plasmid DNA.

Note: See user's manual or package insert for limited label license, and trademark information. For Research Use Only. Not for use in diagnostics procedures.

Anza™ 24 MssI (Invitrogen™)

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3'  C  A  A  A ↑T  T  T  G   5' 

Invitrogen™ Anza™ 24 MssI is a restriction enzyme that cuts DNA at this recognition site: GTTT^AAAC, completely digesting the DNA in 15 minutes at 37°C. Isoschizomers include: PmeI.

For superior convenience, a single buffer and protocol are used with all Anza™ restriction enzymes. The single Anza buffer allows multiple restriction enzymes to be used simultaneously in one reaction. This helps eliminate the need for sequential digests and the DNA clean-up necessary when changing buffers. Additionally, the Anza buffer and restriction enzymes have been designed for flexibility in digestion times. Digestions of up to 16 hours can be carried out without star activity.

Convenient—all Anza enzymes are 100% functional in the single Anza buffer
Fast—complete digestion in 15 minutes
Flexible—overnight digestion without star activity
Simple—a single digestion protocol for all DNA types
Complete system—incorporates Anza DNA Modifying Enzymes for a complete cloning experience

Anza DNA Modifying Enzymes include the Anza T4 DNA Ligase Master Mix, Anza Alkaline Phosphatase Kit, Anza T4 PNK Kit, Anza DNA Blunt End Kit, and Anza DNA End Repair Kit. Use these enzymes for a complete cloning workflow.

Anza Red Buffer, which contains a density gradient and tracking dyes, is supplied for added convenience. Digestion reactions are as efficient in the Anza Red Buffer as in the Anza Buffer. Use of the Anza Red Buffer helps reduce pipetting steps when preparing multiple digestion reactions and a subsequent step includes agarose gel electrophoresis. For applications that require product analysis by fluorescence excitation, the colorless Anza Buffer without the tracking dyes is recommended.

AmpliTaq Gold™ DNA Polymerase, LD (Low DNA) with Gold Buffer and MgCl2 (Applied Biosystems™)

Applied Biosystems® AmpliTaq Gold® DNA Polymerase, LD (Low DNA) is a highly purified version of AmpliTaq Gold® DNA Polymerase that significantly reduces contaminating bacterial DNA sequences commonly present in recombinant protein preparations.

• Manufacturing process results in highly purified enzyme with lower levels of bacterial DNA
• Minimizes non-target, false positive PCR products when amplifying bacterial sequences
• Chemically modified hot-start technique eliminates biological contamination
• GeneAmp® 10X PCR Gold Buffer promotes faster enzyme activation and robust PCR amplification
• Enhanced specificity with time-release PCR is ideal for low-copy target amplification
• AmpliTaq Gold® DNA Polymerase, LD improves both yield and success rate for target amplification

Reliable Results
This highly purified enzyme preparation gives you more confidence in your results and makes advanced PCR techniques quick and easy. When combined with the supplied GeneAmp® 10X PCR Gold Buffer, AmpliTaq® Gold DNA Polymerase, LD provides a flexible hot-start system that is suitable for all PCR applications that require the elimination of bacterial DNA contamination.

Note:
See user's manual or package insert for limited label license and trademark information. For Research Use Only. Not for human or animal therapeutic or diagnostic use.

Ecl136II (EcoICRI) (10 U/µL) (Thermo Scientific™)

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3'  C  T  C ↑G  A  G   5' 

Thermo Scientific Ecl136II (EcoICRI) restriction enzyme recognizes GAG^CTC sites and cuts best at 37°C in its own unique buffer (Isoschizomers: Eco53kI, EcoICRI, Psp124BI, SstI). See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. Note: Also available as a FastDigest enzyme for rapid DNA digestion.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities

Applications

• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP

Note: For methylation sensitivity, refer to product specifications.

DreamTaq Green PCR Master Mix (2X) (Thermo Scientific™)

Thermo Scientific DreamTaq Green PCR Master Mix (2X) is a ready-to-use solution containing DreamTaq DNA Polymerase, optimized DreamTaq Green buffer, MgCl2, and dNTPs. The master mix retains all features of DreamTaq DNA Polymerase. It is capable of robust amplification of up to 6 kb from genomic DNA and up to 20 kb from viral DNA. It ensures higher sensitivity, longer PCR products and higher yields compared to conventional Taq DNA polymerase. Extensive optimization of reaction conditions is not required.

Highlights

• Direct loading of PCR product on gel
• High yields and high sensitivity of PCR
• Amplification of long targets up to 6 kb from genomic DNA and up to 20 kb from viral DNA
• Robust amplification with minimal optimization
• Incorporates modified nucleotides, but does not incorporate dUTP

Applications

• Routine PCR amplification of DNA fragments up to 6 kb
• Genotyping
• High throughput PCR

PasI (10 U/µL) (Thermo Scientific™)

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3'  G  G  G  W  C ↑C  C   5' 

Thermo Scientific PasI restriction enzyme recognizes CC^CWGGG sites and cuts best at 55°C in its own unique buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Features

• Superior quality—stringent quality control and industry leading manufacturing process
• Convenient color-coded Five Buffer System
• Includes universal Tango buffer for double-digestions
• BSA premixed in reaction buffers
• Wide selection of restriction endonuclease specificities

Applications

• Molecular cloning
• Restriction site mapping
• Genotyping
• Southern blotting
• Restriction fragment length polymorphism (RFLP)
• SNP

Note: For methylation sensitivity, refer to product specifications.