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 (Fig.1).

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.

T4 DNA Ligase

Intact Genomics T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA.  This enzyme joins DNA fragments with either cohesive or blunt termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
 

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. 

Intact Genomics T4 DNA Ligase displays up to 3-5X higher ligation efficiency than the nearest competitor.

Product Source
E. coli strain expressing a recombinant clone

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. 

Contents & Storage

1. T4 DNA Ligase
2. 10x T4 DNA Ligase Reaction Buffer (w/o ATP)
3. 10 mM ATP

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 T4 DNA Ligase Reaction Buffer (w/o ATP) 
500 mM Tris-HCl, 100 mM MgCl, 100  mM DTT, pH 7.5 @ 25 °C

Note
10x T4 DNA ligase buffer does not contain ATP. You need to add ATP separately.

Unit Definition 
One Weiss unit is defined as the amount of enzyme required to convert 1 nmol of 32P from pyrophosphate into Norit-absorbance material in 20 minutes under standard assay conditions.

Protocol

1. Set up reaction buffer in a microcentrifuge tube on ice. Use a molar ratio of 1:3 vector to insert DNA.

1. Gently mix the reaction and centrifuge briefly.
2. For cohesive ends, incubate 16 °C for overnight or at room temperature for 30 min.
3. For blunt ends, incubate 16 °C for overnight or at room temperature for 2 hrs.
4. Heat inactivate at 70 °C for 15 min.
5. Cool on ice and transform 2 µl of the reaction into 50 µl competent cells.

ig-Fusion Cloning Kit 
Intact Genomics propriety ig-Fusion cloning technology is a simple, rapid and highly efficient cloning kit which allows to directly clone any PCR product(s) to any linearized expression vector at any site. The PCR fragments can be generated by Intact Genomics high fidelity Pfu DNA polymerase or other high-fidelity DNA polymerases, with primers having 15 to 18 bases of homology at their linear ends to where the product need to fuse. The linearized vector can be generated by PCR or restriction enzymes. The kit is so robust that multiple DNA fragments can be assembled simultaneously and cloned into one construct in a single reaction step within short times (usually 10-30 min) with more than 95% cloning efficiency.

Benefits

• Clone any insert at any site within any vector
• Restriction enzyme and phosphatase free system
• Joining multiple large fragments at once
• Precise insertion at a desired orientation
• Rapid and high efficiency with > 95% positive clones

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. 

Contents & Storage

• 5x ig-Fusion enzyme premix: -20 °C
• 2x PCR premix: -20 °C
• High efficiency competent cells: -80 °C
• Recovery medium:4 °C or -20 °C

Protocol
1. Linearize the vector by restriction enzyme digestion or inverse PCR and purify the product with spin column.
2. Design PCR primers for the gene of interest with 15 to 20 bp at 5'-extensions that are complementary to the ends of the linearized vector.
3. Amplify the gene of interest with Intact Genomics 2x PCR premix or any other high-fidelity DNA polymerase. Run the PCR product on an agarose gel to determine the integrity of the PCR product.
4. Purify the PCR product with spin column.
5. Set up the ig-Fusion cloning reaction as follows: Insert and vector molar ratio 3:1 produce the highest number of colonies.  

6.  Mix the reaction mixture thoroughly.
7.  Incubate the reaction mixture at 50 °C for 10-30 min, then place on ice. Number of colonies depend on the incubation time, insert size and number of inserts need to clone.
8.  Use 2.0 µl of the reaction mixture and transform into high efficiency ig 10B chemical or electroporation competent cells (included). To get the maximum number of colonies, we recommend to use ig 10B electrocompetent cells (Cat # 1212).

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.

MagBio HighPrep™ Plant RNA Plus Kit

, Beads

MAXI Flex Tube Kit

MAXI Flex Tubes combine two modes of action: electro-elution of nucleic acid molecules from polyacrylamide or agarose gels and dialysis or buffer exchange of protein molecules. Flex Tubes allow rapid, secure, simple loading and recovery, with high performance as the most convenient, user friendly, electro-elution and dialysis system on the market.

KIT CONTENTS

• Flex Tubes 2/10/30/50/ 100 pieces
• Supporting tray (for electro elution protocol) 1ea. (select kits)
• Floating rack (for dialysis protocol) 1ea. (select kits)
• Information and Protocol Manual 1ea.

SPECIFICATIONS

• Membrane cut-off: 3.5K(11bp), 6-8K(18-24bp), 12-14K(36-42bp), 25K(76bp) or 50K(152bp) MWCO
• Tube volume: 3ml
• Dialysis volume: 0.1-3ml
• Min. sample size for extraction: 20µg
• Max. gel slice: 2cm x 1cm
• Membrane ultra-clean, sulfur and heavy metal free. EDTA treated
• Flex Tube MWCO are in kilo Daltons (K) for proteins and corresponding base pairs (bp) for nucleic acids as indicated in the table below:

APPLICATIONS

• Dialysis, electro-elution or buffer exchange with volumes between 0.1-3ml
• Preparation of protein samples for MALDI-MS
• Sample concentration
• Large-scale protein dialysis, such as antibodies and recombinant protein purification
• Removal of contaminating micro-molecules
• Tissue culture extraction purification
• Removal of salts, surfactants, solvents, and detergents
• Complex formation studies (protein-protein, protein-DNA, and protein-RNA)
• pH and buffer adjustment of sample solutions, protein extraction or cell extraction
• High throughput dialysis
• Peptide dialysis, as small as 10 amino acids
• Virus-particles purification

MIDI Flex Tube Kit

MIDI Flex Tubes combine two modes of action: electro-elution of nucleic acid molecules from polyacrylamide or agarose gels and dialysis or buffer exchange of protein molecules. Flex Tubes allow rapid, secure, simple loading and recovery, with high performance as the most convenient, user friendly, electro-elution and dialysis system on the market.

KIT CONTENTS

• Flex Tubes 2/10/30/50/ 100 pieces
• Supporting tray (for electro elution protocol) 1ea. (select kits)
• Floating rack (for dialysis protocol) 1ea. (select kits)
• Information and Protocol Manual 1ea.

SPECIFICATIONS

• Membrane cut-off: 1K(3bp), 3.5K(11bp) or 6-8K(18-24bp) MWCO
• Tube volume: 800µl
• Dialysis volume: 50-800µl
• Min. sample size for extraction: 0.5µg
• Max. gel slice: 1cm x 0.5cm
• Membrane ultra-clean, sulfur and heavy metal free. EDTA treated
• Flex Tube MWCO are in kilo Daltons (K) for proteins and corresponding base pairs (bp) for nucleic acids as indicated in the table below:

APPLICATIONS

• Dialysis, electro-elution or buffer exchange with volumes between 50-800µl
• Preparation of protein samples for MALDI-MS
• Sample concentration
• Large-scale protein dialysis, such as antibodies and recombinant protein purification
• Removal of contaminating micro-molecules
• Tissue culture extraction purification
• Removal of salts, surfactants, solvents, and detergents
• Complex formation studies (protein-protein, protein-DNA, and protein-RNA)
• pH and buffer adjustment of sample solutions, protein extraction or cell extraction
• High throughput dialysis
• Peptide dialysis, as small as 10 amino acids
• Virus-particles purification.

MEGA Flex Tube Kit

MEGA Flex Tubes enable user friendly dialysis or buffer exchange of protein molecules with rapid, secure and simple loading along with high performance yield recovery.

KIT CONTENTS

• Flex Tubes 2/10/30/50/ 100 pieces
• Floating rack (for dialysis protocol) 1ea. (select kits)
• Information and Protocol Manual 1ea.

SPECIFICATIONS

• Membrane cut-off: 1K, 3.5K, 6-8K or 12-14K MWCO
• Tube volume: 10ml, 15ml or 20ml
• Dialysis volume: 3-20ml
• Membrane ultra-clean, sulfur and heavy metal free. EDTA treated
• Flex Tube MWCO are in kilo Daltons (K) for proteins

APPLICATIONS

• Dialysis or buffer exchange with volumes between 3-20ml
• Preparation of protein samples for MALDI-MS
• Sample concentration
• Large-scale protein dialysis, such as antibodies and recombinant protein purification
• Removal of contaminating micro-molecules
• Tissue culture extraction purification
• Removal of salts, surfactants, solvents, and detergents
• Complex formation studies (protein-protein, protein-DNA, and protein-RNA)
• pH and buffer adjustment of sample solutions, protein extraction or cell extraction
• High throughput dialysis
• Peptide dialysis, as small as 10 amino acids
• Virus-particles purification

MINI Flex Tube Kit

MINI Flex Tubes combine two modes of action: electro-elution of nucleic acid molecules from polyacrylamide or agarose gels and dialysis or buffer exchange of protein molecules. Flex Tubes allow rapid, secure, simple loading and recovery, with high performance as the most convenient, user friendly, electro-elution and dialysis system on the market.

KIT CONTENTS

• Flex Tubes 2/10/30/50/ 100 pieces
• Supporting tray (for electro elution protocol) 1ea. (select kits)
• Floating rack (for dialysis protocol) 1ea. (select kits)
• Information and Protocol Manual 1ea.

SPECIFICATIONS

• Membrane cut-off: 6-8K(18-24bp), 12-14K(36-42bp) or 25K(76bp) MWCO
• Tube volume: 250µl
• Dialysis volume: 10-250µl
• Min. sample size for extraction: 0.5µg
• Max. gel slice: 0.4cm x 1.1cm
• Membrane ultra-clean, sulfur and heavy metal free. EDTA treated
• Flex Tube MWCO are in kilo Daltons (K) for proteins and corresponding base pairs (bp) for nucleic acids as indicated in the table below:

APPLICATIONS

• Dialysis, electro-elution or buffer exchange with volumes between 10-250µl
• Preparation of protein samples for MALDI-MS
• Sample concentration
• Large-scale protein dialysis, such as antibodies and recombinant protein purification
• Removal of contaminating micro-molecules
• Tissue culture extraction purification
• Removal of salts, surfactants, solvents, and detergents
• Complex formation studies (protein-protein, protein-DNA, and protein-RNA)
• pH and buffer adjustment of sample solutions, protein extraction or cell extraction
• High throughput dialysis
• Peptide dialysis, as small as 10 amino acids
• Virus-particles purification

These items have been discontinued by the manufacturer.

See our Extraction and Purification page or contact us for more options.

PrecisionPak Small Organism DNA Kit

All-inclusive kit, from lysis to extraction, including protocol, lysis beads, all buffers and reagents, magnetic extraction beads, and additional reagents to ensure high quality DNA.

The PrecisionPak Small Organism DNA extraction kit contains a combination of lysis beads preloaded in sample tubes and lysis buffer for thorough homogenization and high yield. Magnetic bead technology, coupled with additional reagents such as Proteinase K and RNase ensure very high-quality DNA. The magnetic bead technology is innovatively designed to be faster and more efficient than spin-column methods.

Tough Small Organism DNA Kit available for: Caterpillar, Crayfish, Crickets, Fruit Fly (Drosophila), Honeybee, Mosquito, Shrimp, Ticks, Zebrafish (D. rerio), Xenopus Tissue.

Soft Small Organism DNA Kits available for: Hookworm, Nematode (B. xylophilus), Roundworm (C. elegans), and Zooplankton.   
 

Bullseye 1Kb DNA Ladder, Logic

• Ready to use
• Contains 16 DNA bands: 100bp-10Kb
• 920ng DNA/ 6 uL/loading
• Easy quantification of DNA fragments
• Stable at room temperature
• Supplied with 6x sample loading buffer

Bullseye 1Kb DNA Ladder, Logic contains 16 DNA bands ranging from 100bp to 10Kb, formulated so that each band contains an amount of DNA that correlates logically to its size, allowing the user to estimate both the size and the quantity of specific fragments at a glance. It is particularly useful for protocols such as probe labeling, DNA sequencing and optimizing insert/vector ratio in ligation reactions, where DNA concentration must be taken into consideration.

Size: 1200 uL
Storage: Store at -20°C.
Concentration: 920 ng/6 uL

Loading Buffer Composition:

• 10mM Tris-HCl
• 1mM EDTA (pH 8.0)
• 0.02% Bromophenol blue
• 0.02% Xylene cyanol
• 5% Glycerol

Usage: Add 6 uL of Bullseye 1Kb DNA Ladder, Logic directly to wells designated for markers, which will yield the amount as indicated in the picture. You may use more than 6 uL of ladder, depending on well size and level of dye used to visualize the bands. Calculate the amount for each band according to your loading volume.

This brand is being discontinued.

Need a great DNA Ladders at a great price?

Try our PR1MA™ SmartCheck DNA Ladders!

Bullseye 100bp DNA Ladder RTU (Ready-to-Use)

• A unique combination of PCR products and a number of proprietary plasmids digested with appropriate restriction enzymes to yield 11 fragments, suitable for use as molecular weight standards for agarose gel electrophoresis.
• The DNA includes fragments ranging from 100-1,500 base pairs. The 500 and 1,500 base pair bands have increased intensity to serve as reference points.
• The approximate mass of DNA in each band is provided (0.5 ug a load) for approximating the mass of DNA in comparably intense samples of similar size.

Source:

• PCR products and double-stranded DNA digested with appropriate restriction enzymes, are phenol and equilibrated to 10nM Tris-HCl (pH 8.0) and 10 mM EDTA.

Range: 100-1,500 bp (DL1); 250-10,000 bp (DL5)
Number of bands: 11 (DL1); 13 (DL5)
Concentration: 100 ug / mL
Package: 50 ug / 500 uL
Recommended Load: 5 uL / well
Contains orange G & xylene cyanol FF as tracking dyes. (DL1)
Contains bromophenol blue & xylene cyanol FF as tracking dyes. (DL5)

Storage:

• Store at 25°C for 6 months
• Store at 4°C for 12 months
• Store at -20°C for 24 months