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.

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PR1MA™ Taq DNA Polymerase 1X Master Mix

PR1MA™ DNA polymerase 1X Master Mix saves time and cost by enabling direct PCR amplification of unpurified templates. It contains a recombinant, truncated (lacks 5’ to 3’ exonuclease activity), highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus. The enzyme is thermostable up to 98°C for polymerase chain reaction assays and is provided as a complete reaction master mix consisting of reaction buffer, dNTPs, MgCl2, and loading dye and only requires the addition of primers and DNA template. Once PCR is complete, the reaction products can be loaded directly into an agarose gel for analysis.

• Lacks exonuclease activity
• Thermotolerant up to 98°C
• Inhibitor Resistant
• Ideal for colony PCR, genotyping, and GC-rich templates
• Storage temperature: -20°C 

Important note: Please be sure to use the buffer provided with this product to ensure optimal results.

*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.

Proudly made in the USA! Click here for more made in America products!
 

PR1MA™ T4 Gene 32 Protein

PR1MA™ T4 Gene 32 Protein (T4 gp32) is a single-stranded DNA binding (ssDNA) protein required for E. coli bacteriophage T4 replication. It binds and stabilizes ssDNA structures which facilitates electron microscopic examination, and has also been shown to improve restriction digests, improve T4 DNA polymerase activity, and increase the yield of PCR reactions, including those with long amplicons. 

• Optimal temperature: 37°C
• Heat inactivation: 65°C for 20 minutes
• Storage temperature: -20°C
• 10X T4 gp32 reaction buffer included

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. 
 

Proudly made in the USA! Click here for more made in America products!
 

PR1MA™ Reverse Transcriptase

PR1MA™ reverse transcriptase (RT) is an RNA-dependent DNA polymerase that can be used for complementary DNA (cDNA) synthesis from an RNA template and is ideal for use in molecular amplification assays. PR1MA™ RT is a robust enzyme that works in a broad range of temperatures (40 - 72°C) and has RNase H activity.

• Optimal temperature: 55°C
• Heat inactivation: 75°C for 20 minutes
• Glycerol-free buffer available
• Storage temperature: -20°C (standard buffer)
• 10X Isothermal buffer included

Standard Buffer Composition

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

Important note: Please be sure to use the buffer provided with this product to ensure optimal results.

*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.   
 

Proudly made in the USA! Click here for more made in America products!
 

PR1MA™ Reverse Transcriptase, RNase H -

PR1MA™ reverse transcriptase (RT) is an RNA-dependent DNA polymerase ideal for use in RT-PCR and first-strand synthesis of complementary DNA (cDNA) for generation of cDNA libraries from single-stranded RNA, DNA, or RNA:DNA hybrids.

• Decreased RNase H activity enables longer cDNA synthesis (>5 kb).
• Lacks 3’ to 5’ exonuclease activity
• Optimal temperature at 42°C
• Temperature range: 40 to 50°C
• Heat inactivation: 70°C for 20 minutes
• Storage temperature: -20°C
• 10X reaction buffer included 

Buffer composition

• 50% glycerol
• 50 mM Tris-HCl
• 50 mM KCl
• 1 mM DTT
• 0.1  mM EDTA
• 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.

Proudly made in the USA! Click here for more made in America products!
 

Accuris Fast Extraction PCR Kit

The Accuris Fast Extraction PCR Kit features an exclusive single-tube solution that extracts genomic DNA in just 8 minutes. This is swiftly followed by PCR amplification utilizing our High Fidelity Hot Start Red Dye Master Mix, designed for superior endpoint PCR performance. High Fidelity Hot Start Red Dye Master Mix is a 2X formula that incorporates a non-reactive red dye, enabling direct gel loading.

This kit offers versatility to suit different workflows: the PCR-prepared genomic DNA can be directly used in endpoint PCR or can be applied in real-time PCR using either SYBR Green or TaqMan Probe chemistries.

• Offers exceptional versatility and can be used to extract and amplify DNA from a wide range of sample types
• Compatible with animal tissue, plant, saliva, & bacterial samples
• Extraction & Amplification in less than 60 minutes
• Includes High Fidelity Hot Start Red Dye Master Mix to prevent the risk of nonspecific amplification & to visualize bands easily
• Process extracted sample via PCR with included mastermix
• Alternatively, amplify via qPCR (qPCR mastermix not included) 

Each 100 reaction kit includes:

1 tube of High Fidelity Hot Start Red Dye Master Mix (1.25 mL)

1 tube of Fast Extraction Lysis Solution (1.25 mL)

Applications: Extraction & Amplification of Tissue, Plant, Saliva, & Bacteria
 

Proudly made in the USA! Click here for more made in America products!
 

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.
 

Looking for other alternatives?

Visit our PCR Reagents page for more options! 

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