Shop All PCR & Cloning Enzymes

PacI (10 U/µL) (Thermo Scientific™)

5'  T  T  A  A  T ↓T  A  A   3' 
3'  A  A  T ↑T  A  A  T  T   5' 

Thermo Scientific PacI restriction enzyme recognizes TTAAT^TAA sites and cuts best at 37°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. 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.

SgeI (3 U/µL) (Thermo Scientific™)

5'  Cm5N  N  G  N9 3'
3'  G  N  N  C  N13 5'
SgeI cleaves DNA targets containing 5-methylcytosine on one or both DNA strands

Thermo Scientific SgeI restriction enzyme recognizes m5CNNG(9/13)^ sites and cuts best at 37°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: DNA methylated by Dcm or CpG methyltransferases will be a substrate for SgeI. Greater than 3-fold overdigestion with SgeI may result in incomplete cleavage. At least two copies of SgeI recognition sequence are required for an efficient cleavage. Amount of the enzyme required for complete digestion of methylated DNA depends on the number of SgeI recognition sites. DNA cleavage products generated by target site cleavage facilitate the nonspecific cleavage by SgeI. Therefore, optimization of the enzyme amount is recommended for DNA cleavage. pBR322 DNA isolated from E. coli dcm+ strain (#SD0041) can be used as a DNA cleavage efficiency control. SgeI cleaves all six dcm methylated targets on pBR322 DNA. For methylation sensitivity, refer to product specifications.

RNase A (Thermo Scientific™)

Thermo Scientific GeneJET RNase A Solution is a component of the GeneJET Plasmid Miniprep Kit (K0502/K0503) and may be purchased separately.

FastDigest PacI (Thermo Scientific™)

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3'  A  A  T ↑T  A  A  T  T   5' 

Thermo Scientific FastDigest PacI restriction enzyme recognizes TTAAT^TAA site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer.

Thermo Scientific FastDigest PacI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

• 100% activity of all FastDigest enzymes in the universal buffer
• 100% buffer compatibility with downstream applications
• Complete digestion in 5–15 minutes
• Direct loading on gels
• No star activity
• 176 FastDigest enzymes available

Applications

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

Note: The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity, refer to product specifications.

ThermoPrime Taq DNA Polymerase, with 10x buffer and separate vial of 25 mM MgCl2 (Thermo Scientific™)

Thermo Scientific ThermoPrime Taq Taq Polymerase is a 94 kDa recombinant thermostable DNA polymerase obtained by high level expression of the Taq DNA polymerase gene in E. coli. A proprietary buffer component protects ThermoPrime DNA polymerase during high temperature steps, ensuring optimal performance throughout the reaction, leading to increased PCR yields. Convenient Master Mix format is also available for easier reaction set-up.

Highlights

• Enhanced enzyme stability for increased yield
• Advanced buffering system minimizes PCR optimization steps

Applications

• Amplification of fragments up to 5 kb
• Routine PCR

Related Products
ThermoPrime Taq DNA Polymerase, with 10x buffer and separate vial of 25 mM MgCl2
ThermoPrime 2x PCR Master Mix, with 1.5 mM MgCl2

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.

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.

FastDigest PspFI (Thermo Scientific™)

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3'  G ↑G  G  T  C  G   5' 

Thermo Scientific FastDigest PspFI restriction enzyme recognizes CCCAGC(-1/-5)^ site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer.

Thermo Scientific FastDigest PspFI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

• 100% activity of all FastDigest enzymes in the universal buffer
• 100% buffer compatibility with downstream applications
• Complete digestion in 5–15 minutes
• Direct loading on gels
• No star activity
• 176 FastDigest enzymes available

Applications

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

Note: The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity, refer to product specifications.

FastDigest HindIII (Thermo Scientific™)

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3'  T  T  C  G  A ↑A   5' 

Thermo Scientific FastDigest HindIII restriction enzyme recognizes A^AGCTT site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer.

Thermo Scientific FastDigest HindIII is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

• 100% activity of all FastDigest enzymes in the universal buffer
• 100% buffer compatibility with downstream applications
• Complete digestion in 5–15 minutes
• Direct loading on gels
• No star activity
• 176 FastDigest enzymes available

Applications

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

Note: The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity, refer to product specifications.

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.

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.

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.

FastDigest PfeI (Thermo Scientific™)

5'  G ↓A  W  T  C   3' 
3'  C  T  W  A ↑G   5' 

Thermo Scientific FastDigest PfeI restriction enzyme recognizes G^AWTC site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: TfiI.

Thermo Scientific FastDigest PfeI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

• 100% activity of all FastDigest enzymes in the universal buffer
• 100% buffer compatibility with downstream applications
• Complete digestion in 5–15 minutes
• Direct loading on gels
• No star activity
• 176 FastDigest enzymes available

Applications

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

Note: The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity, refer to product specifications.

AmpliTaq™ DNA Polymerase with Buffer I (5,000 units/tube) (Applied Biosystems™)

AmpliTaq® DNA Polymerase is a 94 kDa, thermostable, recombinant DNA polymerase obtained by expression of a modified form of the Thermus aquaticus (Taq) DNA polymerase gene in E. coli. It is the most thoroughly characterized enzyme available for the PCR process and its recombinant nature and purification method provide unparalleled purity and reproducibility, vial-to-vial, lot-to-lot.

Features of this enzyme:
  • AmpliTaq® DNA Polymerase is the most thoroughly characterized enzyme available for PCR, a testimony to its overall utility and efficacy
  • Its thermal activity profile makes it reliable for PCR applications
  • It is QC-tested to guarantee reproducible results

Reliable and robust PCR
The thermal activity profile of AmpliTaq® DNA Polymerase is good for PCR applications because its optimal activity is in the same range at which stringent annealing of primers occurs (55°C–75°C). The enzyme's half-life is ~40 minutes at 95°C, providing thermostability that meets the requirements of most difficult PCR applications. AmpliTaq® DNA Polymerase is supplied with GeneAmp® 10X PCR Buffer I. It is also available with GeneAmp® 10X PCR Buffer II and MgCl2 Solution.

RNase H (5 U/µL) (Thermo Scientific™)

Thermo Scientific Ribonuclease H (RNase H) specifically degrades the RNA strand in RNA-DNA hybrids. It does not hydrolyze the phosphodiester bonds within single-stranded and double-stranded DNA and RNA.

Applications

• Removal of mRNA prior to synthesis of second strand cDNA
RT-PCR and qRT-PCR: removal of RNA after first strand cDNA synthesis
• Removal of the poly(A) sequences of mRNA after hybridization with oligo(dT)
• Site-specific cleavage of RNA
• Studies of in vitro polyadenylation reaction products