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.

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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. 
 

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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.   
 

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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
 

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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.
 

2x Master Mix Kit (1.5 mM MgCl2) 

Bullseye Taq DNA Polymerase Mix is a ready-to-use 2x reaction mix. Simply add primers, template, and water to successfully carry out primer extensions and other molecular biology applications.

Bullseye Taq polymerase, the NH4+ buffer system, dNTPs, and magnesium chloride are conveniently present in the Taq DNA Polymerase Mix. (Inert Red Dye is present in BE180303 only)

Bullseye Taq DNA Polymerase Mix offers several advantages. Set up time is significantly reduced. The chance of contaminating component stocks is eliminated. Reduction of reagent handling steps leads to better reproducibility. Standard tests can be set up with the confidence that results will be consistent every time.

Composition of 2x Taq Master Mix

150 mM Tris-HCl pH 8.5, 40 mM (NH4)2S04, 3 mM MgCl2, 0.2% Tween 20Ò

4 mM dNTPs

2 units/µL AS ONE Taq polymerase

Inert Red Dye & Stabilizer (BE180303 only)

This brand is being discontinued and will only be available while supplies last.

Need a great DNA Safe Stain at a great price?
Try PR1MA! Click here to order.

Bullseye DNA SafeStain,10,000x 

• Safest DNA stain by far
• Replaces EthBr in agarose gel electrophoresis
• Detection of DNA and RNA
• Excitation at 290 nm & 490 nm
• Emission at 530 nm

DNA SafeStain is a new and safe nucleic acid stain for visualization of double-stranded DNA, single-stranded DNA, and RNA in agarose gels. The dyes are developed to replace toxic ethidium bromide (EthBr, a potent mutagen), commonly used in gel electrophoresis for visualization of nucleic acids in agarose gels. DNA SafeStain is non-carcinogenic by the Ames-test. The results are negative in both the mouse marrow chromophilous erthrocyte micronucleus and mouse spermary spermatocyte chromosomal aberration tests.

DNA SafeStain emits green fluorescence when bound to DNA. It has two excitation wavelength peaks when bound to nucleic acid, at 290nm and 490nm and the emission can be detected at 530nm.

Storage:
Store under dark at 4°C or room temperature.

User Instruction: 
DNA SafeStain can be used in the exact same way as EtBr in agarose gel electrophoresis. For example, add 100µl to 1 liter of 1X running buffer (1xTAE or 1x TBE), and use this DNA SafeStain containing buffer to make the agarose gel and run the gel.

Special Notes: 
1. For best results, dilute the DNA SafeStain 1:10,000 in your gel running buffer and use this dye-containing running buffer to make your agarose gel and to run the gel.

2. As is the case with EtBr; add more DNA SafeStain, if brighter DNA bands are desired. To view the results, any conventional DNA gel viewing light box or gel documentation system should work well with the DNA SafeStain; although, using a LED light instead of a UV light source is preferred due to the harmful nature of UV light.

This Product is For Research Use Only

*For the proper disposal of this product, follow University or Company Guidelines.

Bullseye RT 2X Master Mix

• Up to 9kb cDNA synthesis
• Ensures sample to sample consistency
• Large RNA sample volume capacity
• Ready to use

Size: 100 rxns

RT 2X Master Mix is a proprietary, ready-to-use master mix for first-strand cDNA synthesis in a 2X concentration. This optimized reaction mix contains ribonuclease inhibitor, dNTPs, and a balanced concentration for oligo(dT) and random primers. The ribonuclease inhibitor effectively protects RNA template from degradation. The oligo(dT) anneals selectively to the poly(A) tail of mRNAs and the random primers do not require the presence of poly(A) and they are utilized for the transcription of mRNA 5-end regions. The resultant cDNA can be directly used as template in different PCR experiments.

Kit Components

Storage

• Store at -20°C in a frost-free freezer.

Application

• cDNA synthesis
• Construction of cDNA libraries
• Generation of probes for hybridization 

Protocol

1. Thaw RNA templates and all reagents on ice. Mix each solution by vortexing.
2. Assemble the following components in a tube on ice, and mix well:

1. Heat the mixture at 65°C for 5 mins and incubate on ice for at least 1 min.
2. Collect all components by a brief centrifugation and add 1 µl of the EasyScript RTase to the tube.
3. Mix well and collect all the components by a brief centrifugation.
4. Incubate the tube at room temperature for 10 min for annealing.
5. Perform cDNA synthesis by incubating the tube for 50 min at 42°C.
6. Stop the reaction by heating it at 85°C for 5 min.
7. Chill on ice. The newly synthesized first-strand cDNA is ready for immediate downstream applications. 

Bullseye EasyScript cDNA Synthesis Kit

Application

• First strand cDNA synthesis for PCR
• Construction of cDNA libraries
• Generation of probes for hybridization

EasyScript cDNA Synthesis Kit is a complete system for the efficient synthesis of first strand cDNA from RNA templates. The recombinant RNasin Ribonuclease Inhibitor, supplied with the kit effectively protects RNA template from degradation.

The kit is also supplied with both oligo(dT) and random primers. The oligo(dT) anneals selectively on the poly(A) tail of mRNA. Random primers do not require the presence of poly(A). Therefore, they can be used for transcription of the 5'-end regions of mRNA. Gene-specific primers may also be used with the kit. The first strand of cDNA can be directly used as a template in PCR.

EasyScript Reverse Transcriptase (RTase) within the kit is a genetically modified form of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV). Both the EasyScript and EasyScript Plus are RNase H deficient (negative). The enzyme is purified from bacteria host as a single holoenzyme of 71 kDa with the capacity of first-strand cDNA synthesis of template up to 9kb and can be used for routine cDNA synthesis.

Kit Components

Storage Buffer
 
50 mM Tris-HCl (pH 8.3), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.

Storage
 
Store at -20°C in a frost-free freezer. Multiple freezing and thawing of RNA should be avoided. Keep RNA on ice all the time. It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated.

General Protocol

RT-PCR reactions should be assembled in a RNA-free environment. The use of "clean", automatic pipettes designated for PCR and aerosol resistant barrier tips are recommended. 
1. Thaw template RNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes.
2. Prepare the following reaction mixture in a PCR tube on ice:

3. Incubate at 25°C for 10 minutes if random primer is used. Omit this step if Oligo(dT) primer or sequence specific primer are used.
4. Incubate the mixture at 42°C for 60 minutes.
5. Stop the reaction by heating at 85°C for 5 minutes.
6. Chill on ice. The newly synthesized first-strand cDNA now can be used directly for PCR amplification.

Notes
 
1. Isolation of poly(A)+ RNA from total RNA is not mandatory; however, doing so may improve the yield and purity of the final product.
2. RNA sample must be free of contaminating genomic DNA.
3. Unlike the oligo(dT) priming, which usually requires no optimization, the ratio of a random primer to RNA is critical in terms of the average length of cDNA synthesized in the reaction. Increasing the ratio of random primer/RNA will result in higher yield of shorter (~500 bp) cDNA, whereas decreasing this ratio will produce longer products.

4. The synthesized cDNA should be stored at -20°C.