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

New and Improved! Ideal for General PCR, Genomic analysis and TA cloning!

Bulleye 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
• This product is not recommended for work with neo-primers

Storage: -20°C

Quality Control:
Taq DNA Polymerase is highly purified, free of contaminating endonucleases, exonucleases and nicking activity. For endonuclease assay, 1µg 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 10nmoles of 4 radioactive labeled dNTPs into acid-insoluble material in 30 minutes at 74°C.

Storage Buffer:
5 units/µl in 50mM Tris-HCl (pH8.0), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.5% TritonX-100, and 0.5% NP-40.
10x Reaction Buffer 100mM KCl, 100mM Tris HCl (pH9.0), 80mM (NH4)2SO4, and 1.0% Triton X-100. (Mg++free): is optimized for use with 200µM dNTPs.

Magnesium Chloride:
25mM MgCl2: In general, 1.5mM MgCl2 is recommended; this may vary with different conditions and primer sets.

Please give us a call for a sample.

For laboratory research only. Not for clinical applications.

• 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 25mM 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, 1.5mM 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 50µl reaction, use 25µl of the Taq Master Mix, add template, primers, and water to a final volume of 50µl. 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 minutes/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: 10mM KCl, 20mM Tris HCl (pH9.0), 16mM (NH4)2SO4, 0.1% Triton X-100, 1.5mM MgCl2, 200mM dNTPs, 2.5units/25ul of Taq DNA polymerase, (optional: trace amount of red or purple dye) and enzyme stabilizers.

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

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

AlumaSeal® Easy-to-pierce Aluminum Foil Seal for Direct Sample Recovery
 
AlumaSeal® 384 provides a secure seal for 384-well plates to minimize evaporation, reduce cross-contamination and prevent spillage.

AlumaSeal® 96 provides a secure seal for 96-well raised-rim and non-skirted PCR plates to minimize evaporation, reduce cross-contamination and prevent spillage. 

• Certified DNase- & RNase- free
• Ideal for use with 384-well and deep-well plates (or select AlumaSeal® 96™ for smaller cut for raised-rim and non-skirted PCR plates.)
• Features one end-tab with no perforation
• Pierceable with a pipet tip or robotic probe for direct sample recovery
• Large cut allows for easier positioning of the foil to the plate via edge of plate alignment vs. centering
• Available in roll format for automation: AlumaSeal® Roll-Seal

• PCR
• Long-term storage
• Light sensitive assays
• Robotics

• -80°C to +120°C

• End-tab removal before thermal cycling is highly recommended
• Remove diagonally from a corner to prevent tearing
• Selection of the proper film size will minimize evaporation
• The film should not overhang or ride up the wall of a raised-rim plate
• More complete coverage of skirted flat-top plates will reduce evaporation in the outer wells

PRIME G (gradient) Thermal Cycler

• Colour touchscreen for fast program setup
• Small space-saving footprint
• Temp Range 4°C to 100°C
• Fast ramp rate of up to 3.0°C/sec
• Larger block sizes; 48 x 0.2ml, 30 x 0.5ml and half a 96 well plate in horizontal format.
• Gradient can be applied across a Temp Range of 30°C to 80°C
• Maximum Gradient 30°C
• Gradient calculator function
• Data transfer via USB
• 4 year warranty

The 3 PRIME G is a small gradient thermal cycler that builds on all the features of the 3 PRIME X instrument. The 48 well block format offers eight columns for annealing temperature optimisation and six rows for optimising reagents such as MgCl2 and primer concentrations. Annealing temperatures can be optimised over 30oC between temperatures 30°C to 80°C. The gradient calculator function displays the temperature for each of the eight columns, ensuring easy replication of thermal conditions.

Technical Specification

Please know that the manufacturer has suspended manufacturing this product indefinitely. 

Contact your trusted MIDSCI Lab Consultant
or email tech@midsci.com for more information.

Key benefits
Speed

• 40 cycles in 40 minutes before fast optimization.
  
• 40 cycles in 15 minutes when optimized.
  
• Genotype in 4 minutes with over 99% accuracy.
  
• Set up a 4plex reaction in only 4 mouse clicks.
  
  Confidence
  
• MIQE compliant - always consistent, always compliant.
  
• Instant settle time to achieve world leading uniformity of ±0.1°C across the block - any well, any instrument - consistent results in any system.
  
• 400 specific qPCR reagent kits available.
  
  
  Performance
  
• Consistent results provide trust and confidence. Well-to-well, run-to-run , machine-to-machine.
  
• Compile data from multiple experiments. Pool plates and results across labs and instruments.
  
• Record every well, with every filter, in every cycle. Never miss a data point.
  
• Adaptive LED Control means you never have uncertainty of saturated unknowns, no bleaching though light to neighboring wells or maximizing the wide dynamic range. Trust results are correct. 
  
• All other block-based qPCR machines have lower uniformity. If you perform Next Generation Sequencing, any other instrument than Prime Pro 48 is risking your NGS, with significant cost implications.
  
  Value
  
• No other qPCR system offers so much performance for such a low price.
  
• Available as a complete system - Techne instrument, plates, seals and reagents.
  
• Alternatively, is chemically open platform - use your preferred supplier reagents.
  
• Lowest price per run - use duplicates and low sample volumes. Don't waste money with triplicates and large sample volumes.
  
• Everything as standard - no add-ons, HRM as standard, unlimited software license. 
  
• Highly accurate and uniform qPCR prevents means successful NGS runs and significant moneysaving.
  
  Sensitivity
  
• ±0.1°C uniformity across the whole block instantly after every temperature change means that any well, in any instrument, provides same answer
  
• HRM - Identify Class IV SNP >99.9% of the time
  
• Unsurpassed block uniformity with patented block technology enables the researcher to use duplicates on Prime Pro 48, whereas triplicates are required on less uniform competitor machines.

   

  The Prime Pro 48 real time PCR system is a new, high specification, economically priced real time thermal cycler from Techne. Made in the UK, the Prime Pro 48 features the highest thermal block uniformity and fastest run time of any block-based qPCR system on the market.
  
  High thermal uniformity and validated sample volumes down to 5ul ensures Prime Pro 48 allows researchers to bring run times for an optimized 40 cycle reaction down to only 15 minutes. PCR efficiency relies on precise temperature control during the denaturation and annealing steps. For the highest accuracy, temperature must remain uniform across the plate, ensuring that all samples are processed equally. The unique thermal block design of Prime Pro 48 achieves this with a
  unique heating and cooling system that quickly cycles from one temperature to the next and then achieves a uniform ±0.1°C in every well of the block within a fraction of a second of reaching each dwell temperature. No other block-based qPCR system can achieve similar performance.
  
  The capacity of Prime Pro 48 accommodates a unique and economical 48-well PCR plate, the size of which is only 1/8 the size of a standard 96-well plate. The well format mirrors a 384-well plate therefore allowing the use of a 16-channel micropipette. The small plate cuts reagent costs in half whilst still producing a strong fluorescence signal. Minimizing the plate size also significantly improves thermal uniformity.
  
  Technical Specification

  High Resolution Melt	Yes
  Volume per well	Validated for 5 to 20 ul
  Detection sensitivity	1 copy
  Temp uniformity	+- 0.1 °C within 1 sec
  Temp stability	+- 0.1 °C
  Temperature range	30 to 100 °C
  Average ramp rate	5.5 °C sec
  Thermal system	Hollow silver block peltier-based with conductive fluid
  Block format	48-well block
  Consumables	48-well custom plates and optical adhesive seals
  Optical system	Dual LED excitation (452 to 486 nm and 542 to 582 nm)
  Camera	CCD camera
  Filters	4 emission filters (505 to 545nm 562 to 596nm 604 to 644nm 665 to 705nm)
  Calibrated dyes	SYBR FAM HEX ROX Cy5
  Additional dyes	All dyes within range compatible with no additional calibration
  Passive reference dyes	Use of ROX is supported but optional
  Data collection	Data always collected in all four filters for all wells
  Plate setup	Plate setup for data analysis can be altered after run completes
  Melt curve	Supports continuous data acquisition in a single filter to provide increased data point collection and reduced run times
  PCR cycle time	40 cycles in less than 40 minutes
  PCR cycle time (FAST optimized)	40 cycles in less than 20 minutes
  Dynamic range	Greater than 9 logs
  Calibration	Not required
  Installation	Plug and play design
  Precision	Discriminates 5000 and 10000 template copies with 99 percent confidence
  Warranty	1 year (parts and labor included)
  Voltage	120 to 240 V
  Frequency	50 and 60 Hz
  Nominal current draw	8A
  Peak power	500W (typical power is 180W)
  Software	Multiple license software included at no additional cost
  Chemistry	All chemistries supported
  Applications	Absolute Quantification and Relative Quantification
  Applications	Allelic Discrimination and High Resolution Melt
  Dimensions closed (WxDxH)	34.5cm x 31cm x 32cm (13.6 in x 12.2 in x 12.6 inches)
  Dimensions open (WxDxH)	34.5cm x 31cm x 37cm (13.6 in x 12.2 in x 14.5 inches)
  Weight	13.6 Kg (30 lb)

   

   

  Ordering information

  Part number	Description
  PRIMEPRO48	Prime Pro 48 Real Time PCR System, includes Accessories and Software (Pro and ProControl). PC not supplied
  PROPLATE48	Pack of 50 Real Time PCR Plates, 48-wells
  PROSEAL48	Pack of 50 Real Time PCR Plate seals

PRIME Base Thermal Cycler

• Colour touchscreen for fast program setup.
• Temp Range 4°C to 100°C
• Fast ramp rate of up to 3.4°C/sec
• Can be upgraded for gradient cycling
• Data transfer via USB
• 4 year warranty

This full sized thermal cycler delivers both high performance and high throughput to provide maximum flexibility when processing a large number of samples in parallel. 


User friendly programming is achieved via a colour touchscreen and intuitive software that is standardised across the entire Techne thermal cycler range. A USB port enables the transfer of programmes between instruments and temperature logs to your PC. A real time graphical display provides an instant visualisation of the program status. 

Gradient cycling can be achieved directly using the PRIME G instrument or by upgrading the PRIME BASE unit. This enables the laboratory to manage changing cycling requirements in a cost effective manner.

The Techne PRIME cycler range is one of the most affordable full sized thermal cyclers on the market and is backed up by a 4 year warranty.

Technical Specification

Quick View

5 Prime G gradient thermal cycler

Please know that the manufacturer has suspended manufacturing this product indefinitely. 
 
Contact your trusted MIDSCI Lab Consultant
 or email tech@midsci.com for more information.

PRIME G (gradient) Thermal Cycler

• Colour touchscreen for fast program setup.
• Versatile block format
• Temperature range 4°C to 100°C
• Fast ramp rate of up to 3.4°C/sec
• Gradient can be applied across a temperature range 30°C to 80°C
• Maximum Gradient 30°C
• Gradient calculator function
• Data transfer via USB
• 4 year warranty

The PRIME G is a gradient enabled thermal cycler with all the features of the PRIME BASE unit. The wide linear gradient with a range of 30°C allows annealing temperatures to be optimised in one experiment. The gradient calculator function displays the temperature for each of the eight columns, ensuring easy replication of thermal conditions.

Technical Specification

• Fast ramping - up to 5°C/second
• Intuitive graphical programming
• Lightweight and compact
• Interchangeable blocks - 24 x 0.2 ml and 18 x 0.5 ml
• Economical - half the price of a full size unit
• 18 month warranty