PR1MA™ Taq

• Ideal for routine PCR applications as well as genotyping, colony PCR, and fast PCR
• Improved template affinity and solubility for higher enzyme activity and greater yields
• Proprietary buffer optimized for a variety of assay conditions
• Conveniently supplied as a 2X Master Mix or in a 2-tube format of polymerase and separate 5X buffer

PR1MA™ Taq provides superior results for routine applications. Modified to improve DNA-binding, this polymerase offers higher solubility and greater template affinity, resulting in consistently superior performance. PR1MA™ Taq Polymerase exhibits a 5' to 3' nuclease activity, but no 3' to 5' (proofreading) activity and works well with a wide range of DNA templates including GC-rich sequences.

The polymerase is supplied with a 5X buffer containing MgCl2 and a proprietary mix of enhancers (dNTP's not included). For convenience, PR1MA Taq is also available in a ready-to-use 2X Master Mix - just add primers and template DNA. Our "Red Dye" Master Mix incorporates a red-loading dye that allows amplified samples to be loaded directly on an agarose gel.  The red color aids in visualization during pipetting, and higher density of the buffer ensures that the samples will drop into the gel wells.

• Replaces hazardous Ethidium Bromide (EtBr)
• Better sensitivity than EtBr
• Detect as little as 0.1ng of DNA
• Two types available
• PS Pre Stain
• LD Loading Dye
• Excitation by UV light or blue light
• Compatible with Accuris SmartBlue™ Transilluminator
• Ships at ambient temperature (stored at 4°C)

PR1MA™ SmartStain™ PS (Pre-Stain) can be used as a direct replacement for Ethidium Bromide in agarose and polyacrylamide gel electrophoresis. The stain emits green fluorescence when bound to dsDNA or ssDNA and emits red fluorescence when bound to RNA. PR1MA™ SmartStain™ PS exhibits excitation peaks at 290 nm and 490 nm, allowing it to be used with UV and blue light.

Protocol:

• Prepare 100 mL of agarose or polyacrylamide solution.
• Add 5 µL of PR1MA™ SmartStain™ to the gel solution before pouring gels.
• For enhanced results, add PR1MA™ SmartStain™ PS to the running buffer at a ratio of 5 µL per 100 mL. Adding PR1MA™ SmartStain™ PS to the running buffer will result in increased sensitivity and better detection of small quantities of nucleic acid.
• After electrophoresis is complete, view the gel using a UV or blue light illuminator   

 
PR1MA™ Taq Plus

• Higher fidelity for long PCR amplicons, up to 35 kb
• Ideal for problematic templates
• Better sensitivity and higher activity for low-copy and long PCR
• Enzyme of choice for TA cloning

PR1MA™ Taq Plus, an optimized blend of our Hot Start Taq and a proofreading polymerase, provides 5X better fidelity than wild-type Taq and increased enzyme activity. It is the perfect choice for GC and AT rich templates, low copy number, and long PCR.

PR1MA™ Taq Start Taq DNA Polymerase 

PR1MA™ Taq Plus Master Mix, 2X Concentration

 
PR1MA™ Hot Start Taq

• Exceptional sensitivity for low-copy PCR
• Ideal for multiplex PCR and amplification of GC-rich DNA
• Same enhanced features as PR1MA Taq Polymerase
• Buffer optimized for fast cycling and reproducibility

Engineered for controlled polymerase activity, PR1MA Hot Start Taq is bound with a monoclonal antibody that blocks enzyme activity. This allows reactions to be set up at room temperature without the risk of non-specific amplification.

When samples are ready, the reaction mixture is heated to 95°C to denature the antibody and initiate the reaction. Similar to the standard PR1MA Taq, Hot Start Taq is provided with a 5X buffer, or in a ready-to-use 2X Master Mix. The Master Mix can be ordered with or without an incorporated red gel loading dye.  The Red Dye Master Mix incorporates a red-loading dye that allows amplified samples to be loaded directly on an agarose gel.  The red color aids in visualization during pipetting, and higher density of the buffer ensures that the samples will drop into the gel wells.

 
PR1MA™ High Fidelity Polymerase

• 50X higher fidelity than Taq DNA polymerase
• Works with crude DNA sample
• Ideal for cloning, mutagenesis and microarrays
• Produces blunt end products, to clone directly into blunt end vectors
• Optimized buffer system with unique PCR enhancers

For applications requiring highly accurate amplification, choose PR1MA™ High Fidelity DNA polymerase. Modified for better solubility and higher activity across a broad range of ionic conditions, this polymerase will amplify a wide range of targets, including those that are GC or AT rich as well as crude samples.

A 3'-5' proofreading exonuclease activity and an error rate of 4.55×10-7 makes PR1MA™ High Fidelity DNA Polymerase the perfect partner for cloning applications. The supplied 5X buffer with dNTPs is optimized for compatibility with a variety of targets.

 

Specifications

 
PR1MA™ High Fidelity Hot Start Master Mix

• Leaves an A-overhang for TA cloning
• Ideal for difficult, high GC content sequences
• 10x fidelity of native Taq
• Ideal for long PCR, up to 10kb targets

PR1MA™ High Fidelity Hot Start Mix is a hot start 2x formulation which provides excellent sensitivity in low-copy number assays with 10x higher fidelity than Taq polymerase. The 2x master mix contains proprietary enhancers, an antibody-mediated hot-start mechanism, and a proofreading component for trouble-free PCR reaction assembly and performance.

The pre-optimized Hot Start Master Mix is supplied in a single tube, reducing the number of pipetting steps while improving throughput and reproducibility. The highly efficient buffer formulation and hot-start blend provide the ideal conditions for high-performance PCR and inactivity at room temperature thereby eliminating non-specific amplification.

PR1MA High Fidelity Master Mix

• Leaves a blunt end
• Rapid extension: up to 1 kb per 15 seconds
• 100x fidelity of native Taq
• Ideal for shorter, less complex targets

A 2x formulation which provides extreme sensitivity in low copy number assays with 100x the fidelity of native Taq, PR1MA High Fidelity Master Mix is perfect for shorter, less complex targets. The 2x master-mix contains proprietary enhancers and a proof-reading component for trouble-free PCR reaction assembly and performance. High Fidelity Master Mix delivers a unique balance of PCR sensitivity, high fidelity, versatility, and tolerance to inhibitors.

The pre-optimized Master Mix is supplied in a single tube, reducing the number of pipetting steps while improving throughput and reproducibility. The highly efficient buffer formulation provides the ideal conditions for high-performance PCR.

PR1MA™ Mammalian Genotyping Kit

• DNA extraction and amplification in 1 hour
• Proprietary lysis buffer optimized for ear punches, tail snips and other mammalian tissues
• Single tube - no organic solvents or clean up procedures
• Enhanced PR1MA Hot Start Polymerase included

Traditionally, mammalian genotyping protocols have involved Proteinase K digestion, neutralization, organic extraction and lots of hands on time to get PCR-ready DNA.

The PR1MA Mammalian Genotyping Kit is quick and easy to use - add sample to the proprietary lysis buffer and incubate for 5-10 minutes, then add the deactivation buffer and incubate for 10 minutes. The crude lysate can then be amplified using fast PCR PR1MA Hot Start Taq Master Mix with red loading dye (included). The kit contains everything needed - just add sample and primers.

 
 
PR1MA 1 Hour Mammalian Genotyping Kit 

 
 
 PR1MA Genotyping Hot Start Master Mix, 2X Concentration with Red Dye

Pfu 2x Master Mix 
Pfu DNA Polymerase 2x master mix is ready to use premix which contains Pfu DNA Polymerase, dNTPs, MgCl2 and stabilizers with optimized reaction buffer. It has been optimized for routine PCR applications. Pfu DNA Polymerase is a heat stable DNA polymerase which has 5' 3' DNA polymerase and 3' 5' exonuclease (proofreading) activities. Pfu DNA polymerase retains the high fidelity, sensitivity and processivity with an error rate six-fold lower than Taq DNA polymerase, and significantly lower than the error rates of most other proofreading enzymes or DNA polymerase mixtures. Pfu 2x Master Mix product is supplied with the unique Intact Genomics 5x Magic Enhancer that enables efficient amplification of GC rich templates up to 84%.

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Product Source
E. coli strain expressing a Pfu DNA Polymerase gene from Pyrococcus furiosus.

Contents & Storage

1. Pfu 2x master mix
2. 5x Magic Enhancer

Store all contents at -20 °C.

1x Master Mix Composition 
10 mM Tris-HCl pH 9.0, 50 mM KCl, 1.5 mM MgCl, 0.2 mM dNTPs, 5% Glycerol, 0.08% Igepal CA 630, 0.05% Tween-20, 100 Units/ml Pfu DNA Polymerase.

Protocol
1. Prepare a reaction mix according to the following table:
  

2. Mix the reaction mixture thoroughly.
3. Program the thermal cycler according to the manufacturer's instructions.
4. A typical PCR cycling program is outlined in the following table.

5. Place the PCR tubes in the thermal cycler and start the cycling program.

6. Analyze 5 µl of PCR products by agarose gel electrophoresis. 

Pfu DNA Polymerase 
Pfu DNA polymerase is a heat stable DNA polymerase which has 5' 3' DNA polymerase and 3' 5' exonuclease (proofreading) activities. Pfu DNA polymerase retains the high fidelity, sensitivity and processivity with an error rate six-fold lower than Taq DNA polymerase, and significantly lower than the error rates of most other proofreading enzymes or DNA polymerase mixtures (1). This product is supplied with the unique Intact Genomics 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM, and 5X Magic Enhancer that enables efficient amplification of GC rich templates up to 84%. The physical purity of this enzyme is 98% as assessed by SDS-PAGE with Coomassie® blue staining.

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Product Source
E. coli strain expressing a Pfu DNA Polymerase gene from Pyrococcus furiosus.

Contents & Storage

1. Pfu DNA Polymerase
2. 10x PCR Buffer with Mg²+
3. 5x Magic Enhancer
4. 10 mM dNTP (Cat. # 3312d, 3314d only)

Store all contents at -20 °C.

Storage Buffer 
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C.

10X PCR Buffer with Mg2+ 
100 mM Tris-HCl pH 9.0, 15 mM MgCl, 100 mM KCl, 80 mM (NH4)2SO4, 0.5% Igepal CA 630

Unit Definition 
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTP into acid-insoluble form in 30 minutes at 72 °C.

Protocol
1. Thaw 10x PCR Buffer, dNTP mix, primer solutions, 5x Magic Enhancer (if required) and mix thoroughly before use. 
2. Prepare a reaction mix according to the following table: The reaction mix typically contains all the components needed for PCR except the template DNA.

3. Mix the reaction mixture thoroughly.
4. Add template DNA to the individual PCR tubes containing the reaction mixture. 
5. Program the thermal cycler according to the manufacturerâs instructions. A typical PCR cycling program is outlined in the following table.

Cas9 Nuclease 
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. The CRISPR system consists of a short non-coding guide RNA (sgRNA) made up of a target complementary CRISPR RNA (crRNA) and an auxiliary transactivating crRNA (tracrRNA). The sgRNA guides the Cas9 endonuclease to a specific genomic locus via base pairing between the crRNA sequence and the target sequence, and cleaves the DNA to create a double-strand break. The location of the break is within the target sequence 3 bases from the NGG PAM (Protospacer Adjacent Motif). The PAM sequence, NGG, must follow the targeted region on the opposite strand of the DNA with respect to the region complementary sgRNA sequence

Intact Genomics Cas9 Nuclease is the purified recombinant Streptococcus pyogenes Cas9 enzyme containing a nuclear localization signal (NLS) at the C-terminal for targeting to the nucleus. This enzyme is designed to perform CRISPR/Cas9-mediated genome editing. The physical purity of this enzyme is â¥98% as assessed by SDS-PAGE with Coomassie® blue staining.

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Cas9 nuclease is free from detectable RNase, Endonuclease (nicking) and non-specific DNase activities.

Product Source 
E. coli BL21 (DE3) strain expressing a Cas9 gene from Streptococcus pyogenes with an N-terminal 6xHis tag and C-terminal SV40 nuclear localization signal (NLS).

Contents & Storage

1. Cas9 Nuclease
2. 10x Cas9 Nuclease Reaction Buffer

Store Cas9 Nuclease and Buffer at -20 °C

Storage Buffer 
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C

1x Cas9 Reaction Buffer 
20 mM HEPES, 100 mM NaCl, 5 mM MgCl, 0.1 mM EDTA, pH 6.5 @ 25 °C

Functional Testing
Cas9 Nuclease functional testing was done by in vitro DNA cleavage assay with the following protocol which gives more than 95% digestion of the substrate DNA as determined by agarose gel electrophoresis.
1) Set up 30 µl reaction in a microcentrifuge tube on ice with the following combinations.

2) Gently mix the reaction mixture and centrifuge briefly.
3) Incubate at 37 °C for 60 min. 
4) Add 1 µl RNase (4 mg/ml) 
5) Incubate at 37 °C for 20 min.
6) Run 0.7 to1% agarose TBE gel.