Shop All PCR & Cloning Enzymes

Phusion U Green Multiplex PCR Master Mix (Thermo Scientific™)

Thermo Scientific Phusion U Green Multiplex PCR Master Mix is a ready-to-use, 2X end-point PCR master mix designed for simultaneous amplification of multiple targets in a single tube. Over 20 primer pairs may be combined into a single reaction for highly specific and efficient multiplexing over a broad range of primer and template concentrations.

The Phusion U Green Multiplex PCR Master Mix contains a proprietary buffer with balanced concentrations of all PCR components eliminating the need for tedious optimization. The Green format further simplifies the workflow - it includes a density reagent and two electrophoresis tracking dyes for direct loading of PCR products on gels.

Highlights

• Specific and efficient multiplex PCR of over 20 targets up to 2.5 kb
• Excellent results with templates of suboptimal purity due to inhibitor tolerance
• Fast cycling and direct loading of PCR products on gels

Applications

• Genotyping
• Pathogen detection
• Food testing
• Analysis of genetically modified organisms
• Amplification of microsatellites

The master mix is based on Phusion U Hot Start DNA Polymerase, a high performance enzyme developed using fusion technology. Similarly to other enzymes from Phusion family, Phusion U DNA Polymerase incorporates more nucleotides per binding event than any other DNA polymerase. This allows achieving high yields of PCR products with shorter extention times and consequently reduced total protocol times. Significantly enhanced processivity also enables successful multiplex PCR on difficult targets such as GC-rich templates or templates of suboptimal purity. Being highly tolerant of many PCR inhibitors, Phusion U Multiplex PCR Master Mixes can even be used with unpurified samples such as blood or serum.

Specificity of multiplex PCR is increased by Affibody-mediated hot start mechanism. Reversibly bound Affibody ligand inhibits the polymerase at ambient temperatures preventing amplification of nonspecific products and formation of primer-dimers.

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.

FastDigest BfoI (Thermo Scientific™)

5'  R  G  C  G  C ↓Y   3' 
3'  Y ↑C  G  C  G  R   5' 

Thermo Scientific FastDigest BfoI restriction enzyme recognizes RGCGC^Y site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: HaeII, AccB2I, Bme142I, BsmHI, Bsp143II, Bst16I, Bst1473II, BstH2I, Btu34II, HinHI, LpnI, NgoAI, NgoBI, NgoCI, NgoGI, NgoJI, NgoMI, NgoWI.

Thermo Scientific FastDigest BfoI 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.

Maxima Reverse Transcriptase (200 U/µL) (Thermo Scientific™)

Thermo Scientific Maxima Reverse Transcriptase (RT) was developed through in vitro evolution of M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity as well as RNase H activity. The engineered enzyme features dramatically improved thermostability and robustness, and increased synthesis rates compared to wild type M-MuLV RT.

Maxima Reverse Transcriptase is capable of reproducible cDNA synthesis from a wide range of input total RNA amounts (from 1 pg to 5 µg) at elevated temperatures (50 to 65°C), which makes this enzyme an ideal tool for two step RT-qPCR (see supporting data).

Due to its high thermostability, Maxima enzyme maintains full activity during the entire reverse transcription reaction, generates the highest yields of cDNA, and is able to synthesize even very long RNA transcripts up to 20 kb. The reaction temperature can be increased up to 65°C for efficient transcription of RNA regions with a high secondary structure or to improve specificity using gene specific primers. Due to its increased synthesis rate, the reverse transcriptase reaction can be completed in 15 to 30 min.

Highlights

• High yields of full-length cDNA up to 20 kb
• Active up to 65°C
• Thermostable—90% active after incubation at 50°C for 60 minutes
• High sensitivity—reproducible cDNA synthesis from a wide range of total RNA quantities (1 pg to 5 µg)
• Efficient—complete cDNA synthesis in 15 to 30 minutes
• Incorporates modified nucleotides

Applications

• Two step RT-PCR
• Two step RT-qPCR
• First strand cDNA synthesis
• Construction of full length cDNA libraries
• DNA labeling
• Primer extension

Also available: Maxima First Strand cDNA Synthesis Kit for RT-qPCR

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.

Anza™ 15 XmaJI (Invitrogen™)

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

Invitrogen™ Anza™ 15 XmaJI is a restriction enzyme that cuts DNA at this recognition site: C^CTAGG, completely digesting the DNA in 15 minutes at 37°C. Isoschizomers include: AspA2I, AvrII, BlnI.

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.

KpnI, HC (50 U/µL) (Thermo Scientific™)

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

Thermo Scientific KpnI restriction enzyme recognizes GGTAC^C 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. Isoschizomers: Asp718I.

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.

Anza™ 49 SmiI (Invitrogen™)

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

Invitrogen™ Anza™ 49 SmiI is a restriction enzyme that cuts DNA at this recognition site: ATTT^AAAT, completely digesting the DNA in 15 minutes at 37°C. Isoschizomers include: SwaI.

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.

True Allele™ PCR Premix (Applied Biosystems™)

Applied Biosystems® True Allele® PCR Premix is a PCR master mix that contains AmpliTaq Gold® DNA Polymerase, dNTPs, MgCl2, and buffer. It is intended for use with the ABI PRISM® Linkage Mapping Set. True Allele® PCR Premix has been optimized for strong, specific PCR amplification of microsatellite markers.

Accessory Products:
ABI PRISM® Linkage Mapping Set

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.

FastDigest PacI (Thermo Scientific™)

5'  T  T  A  A  T ↓T  A  A   3' 
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.

Platinum™ SuperFi™ Green DNA Polymerase (Invitrogen™)

Invitrogen™ Platinum™ SuperFi™ Green DNA Polymerase provides Platinum™ SuperFi™ DNA Polymerase and a 5X SuperFi™ Green Buffer. The SuperFi Green Buffer contains a density reagent and two tracking dyes for convenient direct gel loading of PCR products.

Benefits of Platinum SuperFi DNA Polymerase include:
• Exceptional >100X Taq fidelity
• High specificity and increased yields with Platinum hot-start technology
• Robust amplification of difficult-to-amplify targets including those of sub-optimal purity or with ˃65% GC content
• Convenient workflow with room temperature reaction setup and 24 h bench top stability of pre-assembled reactions

Platinum SuperFi DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with trusted Platinum™ hot-start technology for the highest success in PCR. Featuring >100X Taq fidelity, Platinum SuperFi DNA Polymerase is ideally suited for cloning, mutagenesis, and other applications benefiting from supreme sequence accuracy.

Platinum SuperFi DNA Polymerase is engineered with a DNA-binding domain resulting in high processivity and increased resistance to PCR inhibitors. This feature also enables fast-cycling protocols and amplification of long targets. The 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.

Applications of Platinum SuperFi 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

Using Platinum SuperFi Green DNA Polymerase
Annealing rules for Platinum SuperFi DNA Polymerase are different from many common DNA polymerases (such as Taq DNA polymerases). For optimal results, use the Tm calculator on our website: www.thermofisher.com/tmcalculator.

Platinum SuperFi Green DNA Polymerase is supplied with a separate vial of SuperFi™ GC Enhancer designed for difficult and GC-rich templates (˃65% GC). The SuperFi Green buffer that is also included contains a density reagent and two electrophoresis tracking dyes allowing direct loading of PCR products on a gel. The dyes in the green buffer do not interfere with PCR performance and are compatible with downstream applications including DNA sequencing, ligation, and restriction digestion.

Platinum hot-start technology efficiently inhibits DNA polymerase activity at ambient temperatures, allowing convenient reaction setup at room temperature. In addition, pre-assembled PCR reactions may be stored at room temperature for up to 24h prior to the PCR.

More Platinum SuperFi DNA Polymerase products ›

PagI (BspHI) (10 U/µL) (Thermo Scientific™)

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

Thermo Scientific PagI (BspHI) restriction enzyme recognizes T^CATGA sites and cuts best at 37°C in O buffer (isoschizomers: BspHI, CciI, RcaI). 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. PagI 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.

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.

EcoRII (10 U/µL) (Thermo Scientific™)

5'  C  C  W  G  G   3'
3'  G  G  W  C  C ↑ 5'

Thermo Scientific EcoRII restriction enzyme recognizes ^CCWGG sites and cuts best at 37°C in O 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. Isoschizomers: AjnI, BseBI, Bst2UI, BstNI, BstOI, Psp6I, PspGI.

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 sent 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: At least two copies of EcoRII recognition site are required for efficient cleavage. For cleavage of DNA substrates with only one copy of recognition site MvaI, neoschizomer of EcoRII is recommended. EcoRII may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation, or heat the digested DNA in the presence of SDS prior to electrophoresis. Assayed using pBR322 DNA (dcm-). EcoRII is blocked by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.For methylation sensitivity, refer to product specifications.

Proteinase K (50 µg/µL) (Invitrogen™)

This Proteinase K is used in QuantiGene assays and is one of two components in the QuantiGene Sample Processing kits. Proteinase K digests proteins such as ribonucleases that could interfere with the quantitation of RNA using QuantiGene assays.

Anza™ 10-Pack Starter Kit (Invitrogen™)

The Invitrogen™ Anza™ 10-Pack Starter Kit includes the 10 common restriction enzymes, 2 DNA modifying enzymes, and buffers listed below:

• Anza™ 11 EcoRI
• Anza™ 5 BamHI
• Anza™ 1 NotI
• Anza™ 16 HindIII
• Anza™ 12 XbaI
• Anza™ 8 XhoI
• Anza™ 3 BcuI (isoschizomer to SpeI)
• Anza™ 10 DpnI
• Anza™ 19 BglII
• Anza™ 26 Eco32I (isoschizomer to EcoRV)
• Anza™ T4 DNA Ligase Master Mix
• Anza™ Alkaline Phosphatase
• Anza™ Buffer (10X)
• Anza™ Red Buffer (10X)

For a kit with just 5 enzymes, see the Anza™ 5-Pack Starter Kit.

The Anza™ Restriction Enzyme Cloning System employs a single buffer and protocol for all Anza restriction enzymes. The single Anza buffer allows multiple restriction enzymes to be used simultaneously in one reaction. This eliminates 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 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.

The Anza 10-pk Starter Kit is a subset of the Anza Restriction Enzyme Cloning System, unifying the traditional cloning processes. For more information on the system, visit www.thermofisher.com/anza.