PR1MA™ Cod Uracil-DNA Glycosylase

PR1MA™ Cod Uracil-DNA Glycosylase (cUNG) is a recombinant, thermolabile enzyme that removes uracil from DNA. It is ideal for preventing carry over contamination during RNA or DNA amplification reactions that substitute dUTP for dTTP. cUNG is the only commercially available UNG that is completely and irreversibly inactivated by moderate heat treatment, unlike bacterial versions of the enzyme. Cod UNG treatment in combination with targeted pre-amplification using dUTP provides a simple and efficient solution to eliminate carry-over contamination and the generation of false positives and inaccurate quantification.

• Optimal temperature: 37°C
• Heat inactivation: 55°C for 5 minutes
• Enables contamination control in PCR and other amplification methods
• Does not degrade product after inactivation, enabling downstream use of the amplicon
• Storage temperature: -20°C

Buffer composition

• 50% glycerol
• 50 mM Tris-HCl
• 50 mM KCl
• 1 mM DTT
• 0.1  mM EDTA
• 0.1% Tween-20
• pH = 7.5

*These products are intended for research use only, not for therapeutic or diagnostic purposes in humans or animals. The safety and efficacy of these products in diagnostic or other clinical uses has not been established.
 

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.

This product is no longer available, please contact us for other options.
 

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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.

HS Taq Polymerase Master Mix BLUE is a ready to use 2 X master mix. Simply add primers, template and water to successfully carry out primer extensions. The master mix is available in 100, 500, 1000, and 2500 reaction sizes and is available with two different buffer options (HS Buffer 1 and HS Buffer 2)

HS Taq Polymerase is a modified form of Bullseye Taq, activated by heat treatment. This results in higher specificity and greater yields when compared to standard Taq. The added benefit of the HS Master Mix Blue is that it can be loaded directly onto the gel as there is no need to use separate loading dyes for subsequent electrophoresis and visualization. This cuts down on the chance of contaminating the component stocks and leads to better reproducibility.

Composition of 2 X HS Master Mix Blue with Buffer 1
Tris-HCl, pH 8.5, (NH4)2SO4,3 mM, MgCl2, .2 % Tween 20, .4 mM dNTPs, .2 units/uL HS Taq Polymerase, Inert Blue Dye, Stabilizer

Composition of 2 X HS Master Mix Blue with Buffer 2
Tris-Hcl, pH 8.5, Balanced KCl/(NH4)2SO4,3 mM, MgCl2, .2 % Tween 20, .4 mM dNTPD, .2 units/uL HS Taq Polymerase, Inert Blue Dye, Stabilizer

Bullseye Taq DNA Polymerase

Bullseye Taq DNA Polymerase is the most popular thermostable enzyme used in DNA amplification experiments. This high performance Taq DNA polymerase is specifically purified to produce excellent yields with little or no background. Its outstanding activity and unique thermal DNA amplification properties make it one of the best-value DNA polymerases available.

• Robust performance
• Leaves 3' A overhang
• Stable at all storage temperatures
• Ideal for general PCR, genomic analysis and TA cloning

Storage: -20°C

Quality Control
Taq DNA Polymerase is highly purified, free of contaminating endonucleases, exonucleases and nicking activity. For endonuclease assay, 1 ug of Lamda/Hind III DNA is incubated with 20 units of enzyme in assay buffer at 75oC for 16 hours with no visible contaminating activity observed. Also, every lot is tested for its performance consistency.

Unit Definition
One unit incorporates 10n moles of 4 radioactive labeled dNTPs into acid-insoluble material in 30 minutes at 74°C.

Storage Buffer
5 units / uL in 50 mM Tris-HCl (pH 8.0), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol, 0.5% TritonX-100, and 0.5% NP-40.
10x Reaction Buffer 100 mM KCl, 100 mM Tris HCl (pH9.0), 80 mM (NH4)2SO4, and 1.0% Triton X-100. (Mg++free): is optimized for use with 200 uM dNTPs.

Magnesium Chloride
25 mM MgCl2: In general, 1.5 mM MgCl2 is recommended; this may vary with different conditions and primer sets.
 
*For laboratory research only. Not for clinical applications. 

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• Pre-optimized ready-to-use PCR reagents
• Less pipetting to avoid contamination
• Quick and easy to set up
• Direct loading for electrophoresis
• Reproducible results

Users only need to add templates and primers, and water if needed. Extra 25 mM magnesium solution is also provided for additional fine adjustments. The Red Mixes contain a non-hazardous inert red dye. The purple mixes contain a non-hazardous inert purple dye. Completed PCR reactions utilizing the D124-R (red dye) or D124-P (purple dye) can be directly loaded into a well of agarose gel or other gels, for electrophoresis. No extra loading buffer is needed.

Storage: 4°C for up to one month, or -20°C for long term storage.

Magnesium Chloride: In general, 1.5 mM MgCl2 is recommended; this may vary with different conditions and primer sets. Some primers/templates may require adjustments for MgCl2 concentration, which can be achieved as shown below:

Directions for use: For a 50 uL reaction, use 25 uL of the Taq Master Mix, add template, primers, and water to a final volume of 50 uL. Cycling conditions vary for different templates and primers. To start with, try 30 cycles as follows: denature at 94°C for 30 seconds, anneal around 55°C for 30 seconds, and extend at 72°C for 1 minute / kb. After the PCR cycles, extend at 72°C for another five minutes to complete the PCR. Then store the reaction at 4°C.

1x Composition: 10 mM KCl, 20 mM Tris HCl (pH 9.0), 16 mM (NH4)2SO4, 0.1% Triton X-100, 1.5 mM MgCl2, 200 mM dNTPs, 2.5 units / 25 uL of Taq DNA polymerase, (optional: trace amount of red or purple dye) and enzyme stabilizers.
 

This brand is being discontinued, please contact us for other options.
 

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Bullseye GC HS 2x Master Mix I is an all-in-one 2x master mix containing HS DNA polymerase, GC Buffer I, enhancer, dNTPs and MgCl2. Simply mix GC HS 2x Master Mix I with primers, template and water and you are ready to carry out successful primer extensions. HS DNA Polymerase is a modified form of Bullseye Taq DNA polymerase, which is activated by heat treatment. A chemical moiety is attached to the enzyme at the active site, which renders the enzyme inactive at room temperature. Thus, during setup and the first ramp of thermal cycling, the enzyme is not active and misprimed primers are not extended. The result is higher specificity, increased sensitivity and greater yields when compared to standard DNA polymerases.

Key Features

• For amplification of DNA targets with high GC content
  
• Convenient reaction set-up at room temperature
  
• High specificity, sensitivity and product yield
  
• Detection of low abundance targets
  
• Diminished formation of non-specific product

   

  Composition of GC HS 2x Master Mix I
  

• HS DNA Polymerase
  
• Optimized buffer components, 3.0 mM MgCl
  
• dNTPs
  
• Enhancer

This brand is being discontinued, please contact us for other options.
 

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General Description
Bullseye GC HS 2x Master Mix II is an all-in-one 2x master mix containing HS DNA polymerase, GC Buffer II, enhancer, dNTPs and MgCl. Simply mix GC HS 2x Master Mix II with primers,template and water and you are ready to carry out successful primer extensions. HS DNA Polymerase is a modified form of Bullseye Taq DNA polymerase, which is activated by heat treatment. A chemical moiety is attached to the enzyme at the active site, which renders the enzyme inactive at room temperature. Thus, during setup and the first ramp of thermal cycling, the enzyme is not active and misprimed primers are not extended. The result is higher specificity, increased sensitivity and greater yields when compared to standard DNA polymerases.

Key Features

• For amplification of DNA targets with high GC content
  
• Convenient reaction set-up at room temperature
  
• High specificity, sensitivity and product yield
  
• Detection of low abundance targets
  
• Diminished formation of non-specific product

  
  Composition of GC HS 2x Master Mix I
  

• HS DNA Polymerase
  
• Optimized buffer components, 3.0 mM MgCl
  
• dNTPs
  
• Enhancer

  
  Storage and Stability
  The unopened kit is stable at -20°C for 1 year

Combo Includes:
200 μL Taq DNA Polymerase (5 units/μL)
4 x 500 μL vials of dNTP Premix (100 mM each of dATP, dCTP, dGTP, dTTP)

D118-1000

High-performance Taq DNA Polymerase specially purified to produce excellent yields with little or no background. Its outstanding activity and unique thermal DNA amplification properties make it one of the most economic DNA polymerases available.

Special Features

• Robust performance
• Leaves 3'A overhang
• Stable at all storage temperatures
• Best price/quality
• APPLICATIONS
• General PCR
• Genomic analysis
• TA cloning

D116

Ready-to-use, ultra-pure, molecular grade solution made from 100mM each of dATP, dCTP, dGTP, and dTTP stock. The PreMix is designed to reduce "hands-on" time for researchers, thereby lowering the possibility of contamination and error. Furthermore, every lot of dNTP solution is pre-tested for optimal performance. The nucleotides are supplied in sterile, doubly distilled water. Each dNTP has a final concentration of 10mM for a total PreMix dNTP concentration of 40mM.

Special Features

• Ready to use
• Each lot functionally tested
• Purity; at least >99%
• APPLICATIONS
• For RT-PCR, large PCR, and real-time PCR
• Reverse Transcription and mutagenesis

PR1MA™ RNA Controls

Looking for safe assay controls for highly infectious viruses or foreign animal diseases?

MIDSCI™ offers a selection of PR1MA™ RNAs – nuclease-resistant, single-stranded RNAs, suitable as process controls for RNA extraction from various sample matrices. These specially engineered, non-infectious, MS2 phage-like particles protect their contents from degradation by nucleases and can package sequences of up to 1.5 kb from viruses such as SARS-CoV-2, foot-and-mouth disease virus, and human immunodeficiency virus.
 

Amount: Between 1E6 or 1E10 copies (cp)
Concentration: Between 1E7 or 1E10 cp / mL, respectively
Volume: 0.1 and 1 mL respectively
Lysis: 65° C for 5 minutes in the RT step or by standard RNA extraction
Storage temperature: 4°C

Buffer composition

• 10 mM Tris HCl
• 100 mM NaCl
• 1 mM MgCl2
• 0.1% gelatin
• pH = 7.0 

*These products are intended for research use only, not for therapeutic or diagnostic purposes in humans or animals. The safety and efficacy of these products in diagnostic or other clinical uses has not been established.