Shop All PCR & Cloning Enzymes

BplI (5 U/µL) (Thermo Scientific™)

5'  N8G  A  G  N5C  T  C  N13 3'
3'  N13C  T  C  N5G  A  G  N8 5'

Thermo Scientific BplI restriction enzyme recognizes ^(8/13)GAG(N)5CTC(13/8)^ sites and cuts best at 37°C in Tango(+SAM) 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: BplI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05 mM S-adenosylmethionine gives more than a 100-fold increase in BplI activity. Still, complete cleavage of some substrates with BplI is difficult to achieve. BplI concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase. For methylation sensitivity, refer to product specifications.

DNase I Solution (2500 U/mL) (Thermo Scientific™)

Thermo Scientific DNase I removes unwanted DNA from cell lysates to improve protein extraction efficiency.

Features of Thermo Scientific DNase I:

• Degrades and removes unwanted DNA from samples
• Cleaves both single-stranded and double-stranded DNA
• Compatible with Thermo Scientific Pierce Cell Lysis Reagents
• Reduces viscosity of bacterial lysates (protein extracts) to facilitate pipetting

Deoxyribonuclease I (DNase I) is a single, glycosylated polypeptide that degrades unwanted single- and double-stranded DNA. The enzyme works by cleaving DNA into 5' phosphodinucleotide and small oligonucleotide fragments. DNase I is commonly added to cell lysis reagents to remove the viscosity caused by the DNA content in bacterial cell lysates or to remove the DNA templates from RNAs produced by in vitro transcription. This grade of DNase is sufficient for protein work. Use RNase-free DNase for any application requiring the digestion of DNA in which it is crucial to avoid damage to RNA.

General information about the use of DNase I:

• Calcium ions are required for activity of DNase I. Trace amounts of Ca++ may be present at high enough concentration for DNase I to be active, however use of EGTA or calcium-free buffers can reduce DNase I activity to undetectable levels.
• High levels (i.e., 100 mM) of monovalent ions such as Na+ and K+ will decrease DNase I activity
• DNase I is inactivated by heating to 65°C for 10 minutes
• Kunitz unit: 1 Kunitz unit is the amount of enzyme required to cause an increase of 0.001 A260nm/min/mL at 25°C in 0.1M NaOAc, pH 5.0 due to degradation of highly polymerized DNA
• Degradation assay units: 1 unit is defined as the amount of enzyme required to completely degrade 1 µg of plasmid DNA in the 10 minutes at 37°C in 10 mM Tris·HCl, pH 7.5, 50 mM MgCl2, 13 mM CaCl2.

Specifications:

• Quantity: 0.5 mL
• Concentration: ≥ 2500 units/mL
• Unit definition: 1 unit is defined as the amount of enzyme required to produce an increase in absorbance at 260nm of 0.001/min/mL at 25°C of highly polymerized DNA.
• Visual: Clear, colorless liquid, free of insoluble material
• Formulation: DNase I in 10 mM Tris-HCl pH 7.5, 10 mM CaCl2, 10 mM MgCl2

Related Products
DNase I Solution (1 unit/µL), RNase-free

Pierce™ HRV 3C Protease Solution Kit (2 units/µL) (Thermo Scientific™)

Thermo Scientific Pierce Human Rhinovirus (HRV) 3C Protease is a recombinant cysteine protease used to remove fusion tags from proteins with the HRV 3C cleavage sequence and is dual tagged for easy removal from the sample after cleavage.

Features of Thermo Scientific Pierce HRV 3C Protease:

Effective—HRV 3C is designed to remove purification tags from your protein of interest
High activity at 4°C—activity is ≥2000U/mg control protein; one unit will cleave 0.1 mg control protein at 4°C after 16h
Flexible—dual-tagged enzyme is compatible with various columns and buffers

Properties:
• Alternative names: Human Rhinovirus 3C
• Source: Escherichia coli (E.coli)
• Molecular weight: 47.8kD
• Form: Liquid
• Protease type: Cysteine
• Optimal reaction conditions: 50 mM Tris-HCl, 150 mM NaCl, pH 8.0

Pierce HRV 3C Protease is highly specific for the recognition sequence Leu-Glu-Val-Leu-Phe-Gln-↓-Gly-Pro and cleaves after the glutamine residue. The recombinant enzyme is engineered as a fusion protein with glutathione-S-transferase (GST) and polyhistidine (6xHis) tags, so that the protease can be easily removed after cleavage using a variety of solid supports with immobilized glutathione (GSH) or metal chelates, respectively. HRV3C is also active in a variety of commonly used buffers, providing further flexibility in experimental design.

Applications:
• Removal of purification tags from fusion proteins

More Product Data
Choosing a vector and purification method for in vitro protein expression

FastDigest KspAI (Thermo Scientific™)

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

Thermo Scientific FastDigest KspAI restriction enzyme recognizes GTT^AAC site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: HpaI, BsaKI, BseII, BstEZ359I, BstHPI, MwhI, SsrI, Uba1408II.

Thermo Scientific FastDigest KspAI 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

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.

RNase T1 (1000 U/µL) (Thermo Scientific™)

Thermo Scientific RNase T1 is an endoribonuclease that specifically degrades single-stranded RNA at G residues. It cleaves the phosphodiester bond between the 3'-guanylic residue and the 5'-OH residue of adjacent nucleotides with the formation of corresponding intermediate 2', 3'-cyclic phosphate. The reaction products are 3'-GMP and oligonucleotides with a terminal 3'-GMP.

RNase T1 does not require metal ions for activity.

Applications

• Removal of RNA from DNA preparations
• RNA sequencing
• Ribonuclease protection assays. Used in conjunction with RNase A
• Removal of RNA from recombinant protein preparations
• Determination of the level of RNA transcripts synthesized in vitro from DNA templates containing a "G-less cassette"

XhoI (10 U/µL) (Thermo Scientific™)

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

Thermo Scientific XhoI restriction enzyme recognizes C^TCGAG sites and cuts best at 37°C in R 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: PaeR7I, Sfr274I, SlaI, StrI, TliI.

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.

Ambion™ RNase A, affinity purified, 1 mg/mL (Invitrogen™)

Ambion® RNase A is an endonuclease that specifically cleaves 3' of U and C residues. Affinity Purified RNase A is intended for critical applications when the absence of DNase and other nonspecific nuclease activities is essential. Supplied in one tube containing 1 mg (1 mg/mL). Note: This preparation contains RNase B, a carbohydrate isoform of RNase A. RNase A is rigorously tested for contaminating nonspecific endonuclease, exonuclease, and protease activity.

FastDigest MnlI (Thermo Scientific™)

5'  C  C  T  C  N7 3'
3'  G  G  A  G  N6 5'

Thermo Scientific FastDigest MnlI restriction enzyme recognizes CCTC(7/6)^ site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer.

Thermo Scientific FastDigest MnlI 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 BseJI (Thermo Scientific™)

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

Thermo Scientific FastDigest BseJI restriction enzyme recognizes GATNN^NNATC site and cuts best at 65°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: BsaBI, Bse8I.

Thermo Scientific FastDigest BseJI 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.

Acc65I (Asp718I) (10 U/µL) (Thermo Scientific™)

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

Thermo Scientific Acc65I (Asp718I) restriction enzyme recognizes G^GTACC sites and cuts best at 37°C in O buffer (Isoschizomers: Asp718I). 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.

Taq DNA Polymerase, recombinant (Invitrogen™)

Taq DNA Polymerase is a thermostable enzyme that synthesizes DNA from single-stranded templates in the presence of dNTPs and a primer. The enzyme consists of a single polypeptide with a molecular weight of 94 kDa. It has a 5´→3´ DNA polymerase activity and a 5´→3´ exonuclease activity.

With our Taq DNA Polymerase, you get:

• Your choice of recombinant or native enzyme
• Amplification of PCR products up to 5 kb in size
• An enzyme that is licensed and qualified for PCR

Applications
Taq DNA Polymerase is appropriate for use in the amplification of DNA from complex genomic, viral, and plasmid templates, RT-PCR, sequencing ssDNA, and cycle sequencing.

Source
Recombinant enzyme is purified from the cloned Thermus aquaticus DNA polymerase gene expressed in E. coli.

Unit definition
One unit of Taq DNA Polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into DNA in 30 min at 74°C.

For superior PCR performance, we recommend DreamTaq DNA Polymerase.

Anza™ 10 DpnI (Invitrogen™)

5'  G  Am6T  C   3' 
3'  C  T    ↑Am6G   5' 

Invitrogen™ Anza™ 10 DpnI is a restriction enzyme that cuts DNA at this recognition site: Gm6A^TC, completely digesting the DNA in 15 minutes at 37°C. Isoschizomers include: MalI.

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.

Anza™ 125 HpyF10VI (Invitrogen™)

5'  G  C  N  N  N  N  N ↓N  N  G  C   3' 
3'  C  G  N  N ↑N  N  N  N  N  C  G   5' 

Invitrogen™ Anza™ 125 HpyF10VI is a restriction enzyme that cuts DNA at this recognition site: GCNNNNN^NNGC, completely digesting the DNA in 15 minutes at 37°C. Isoschizomers include: BstMWI, MwoI.

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.

FastDigest NotI (Thermo Scientific™)

5'  G  C ↓G  G  C  C  G  C   3' 
3'  C  G  C  C  G  G ↑C  G   5' 

Thermo Scientific FastDigest NotI restriction enzyme recognizes GC^GGCCGC site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: CciNI.

Thermo Scientific FastDigest NotI 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.

RNase A/T1 Mix (Thermo Scientific™)

Thermo Scientific RNase A/T1 Mix combines the RNA degradation activity of both RNase A and RNase T1. The RNase A specifically hydrolyzes RNA at C and U residues; RNase T1 specifically hydrolyzes RNA at G residues.

Highlights

• Higher level of RNA degradation than with RNase A and RNase T1 separately
• Free of DNase activity - It is not necessary to heat the mix before use

Applications

• Removal of RNA from DNA preparations
• Removal of RNA from recombinant protein preparations
• Ribonuclease protection assays