PR1MA™ qMAX One-Step RT-qPCR Kits

• RNA to cDNA to qPCR in one tube
• High purity enzyme formulation for enhanced stability and performance
• PR1MA Hot-Start Taq allows for preparation at room-temperature
• Compatible with all qPCR instruments
• Available for green fluorescence or probe detection

Both One-Step Kits are compatible with standard and fast cycling protocols and provide increased sensitivity, speed and reproducibility for a broad range of samples and targets.  The polymerase mix is available with different levels of ROX reference dye for compatibility with all qPCR instruments.

Two versions of our One-Step qPCR Kits are available:

qMAX™ Green One-Step Kits incorporate our proprietary intercalating dye which exhibits higher fluorescent and lower PCR inhibition than other popular dyes such as SYBR.

qMAX™ Probe One-Step Kits are optimized for use with popular TaqMan, Scorpions, and molecular beacon probes.

*Please note these products ship on dry ice.  Appropriate shipping charges apply unless otherwise noted on a quote.

 
PR1MA™ SmartCheck DNA Ladders

• Ready-to-use formulation includes loading buffer and tracking dye
• 500 µL suitable for 100 lanes (5 µL per lane)
• Higher intensity reference bands
• Ultra pure production allows economical ambient shipping

Technical Details
 

Protocol: Briefly vortex the tube and use 5 µL per lane. Additional loading buffer is not required.

Concentration: 0.1 mg/mL

Tracking dye: bromophenol blue and xylene cyanol

Buffer formulation: 10mM Tris-HCl (pH 8.0), 5mM EDTA, 12.5% glycerol, 0.008% bromophenol blue, 0.008% xylene cyanol

Shipping and Storage: SmartCheck™ DNA Ladders are shipped at ambient temperature. On arrival, store at -20°C for optimum stability and long-term storage up to 15 months.

Production: SmartCheck™ ladders are produced from proprietary plasmids, digested to completion. A multistep chromatography method is used to ensure purity and DNA quality.

Quality Control: Each lot of SmartCheck™ DNA Ladders is tested with a spectrophotometer to confirm DNA concentration, as well as agarose electrophoresis to confirm band sharpness.

Item	Description	Volume
PR4005-100	PR1MA™ SmartCheck™ 50bp DNA Ladder	500 µL / 100 Lanes
PR4005-500	PR1MA™ SmartCheck™ 50bp DNA Ladder	5 x 500 µL / 500 Lanes
PR4010-100	PR1MA™ SmartCheck™ 100bp DNA Ladder	500 µL / 100 Lanes
PR4010-500	PR1MA™ SmartCheck™ 100bp DNA Ladder	5 x 500µL / 500 Lanes
PR4100-100	PR1MA™ SmartCheck™ 1kb DNA Ladder	500 µL / 100 Lanes
PR4100-500	PR1MA™ SmartCheck™ 1kb DNA Ladder	5 x 500 µL / 500 Lanes

  

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

IBI RT-PCR Certified and Nuclease-Free Water is ideal for all applications in a molecular biology lab including PCR, RT-PCR, restriction enzyme
assays, modifying enzyme assays, transfection, cloning, transformation, and all general molecular biology lab procedures.

IBI PCR Grade water is manufactured under stringent conditions. The purification process includes continuous deionization, reverse osmosis,
UV-treatment, 0.2µm filtration, followed by steam sterilization in an autoclave.

• RNA to cDNA to qPCR, in one tube
• High purity enzyme formulation for enhanced stability and performance
• Accuris Hot-Start Taq allows for preparation at room-temperature
• Blue dye facilitates pipetting and visualization in plates
• Available for green fluorescence or probe detection
• Multiplex formulation available for multiple target amplification

Accuris qMAX One-Step RT-qPCR kits

• RNA to cDNA to qPCR, in one tube
• High purity enzyme formulation for enhanced stability and performance
• Accuris Hot-Start Taq allows for preparation at room-temperature
• Blue dye facilitates pipetting and visualization in plates
• Available for green fluorescence or probe detection
• Multiplex formulation available for multiple target amplification
• Bulk pricing available for high throughput labs and kit manufacturers contact us for details

Accuris qMAX™ One Step Kits allow for highly sensitive real time RT-qPCR assays to be performed directly from RNA templates. Workflows are simplified with optimized formulations of ready-to-use 2X qPCR master mix and 20X reverse transcriptase.

Optimized buffer includes powerful RNase inhibitors, and an extremely thermostable MMLV-derived reverse transcriptase enables robust first strand cDNA synthesis. Accuris Hot Start Taq uses an antibody mediated hot start mechanism allowing for sample preparation at room temperature. Only after an initial incubation at 95C will the Taq become active, so non-specific amplification is greatly reduced. An inert blue dye is included in the Taq master mix to help simplify pipetting and reduce errors.

Three versions of our One Step qPCR kits are available:

qMAX Green One Step kits incorporate our proprietary intercalating dye which exhibits higher fluorescent and lower PCR inhibition than other popular dyes such as SYBR.

qMAX Probe One Step kits are optimized for use with popular TaqMan, Scorpions, and molecular beacon probes.

qMAX Probe One Step Multiplex kits are specifically developed and optimized for efficient probe-based detection of multiple targets in a single reaction well. The Multiplex formulation is comprised of a 20x reverse transcriptase and 2x PCR Mix preparation ideally suited for complex RNA samples including low-copy number viral RNA commonly used in the clinical and research laboratory. qMAX Probe One Step Multiplex kits have been designed to overcome the many challenges of multiplex RT-PCR, by addressing important factors such as the balance between magnesium chloride and deoxynucleotide concentrations, the relative Taq Polymerase and reverse transcriptase concentration, and the ionic conditions of the core reaction buffer.

All Accuris One-Step Kits are compatible with standard and fast cycling protocols and provide increased sensitivity, speed, and reproducibility for a broad range of samples and targets. The polymerase mix is available with different levels of ROX reference dye for compatibility with all qPCR instruments.

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.

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