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NZYTech Brochure

Agaroses and DNA Stains

For high standard DNA electrophoresis

Agaroses

Portfolio

Agarose

Definition

Agarose is a neutral polysaccharide extracted from the cell walls of certain Rhodophyceae algae, also known as agarophyte seaweeds because they are the raw material used in the production of agar-agar. The structure of the polysaccharide is a galactan, formed by linking agarobioses 1-3, 1-4, as shown in the above illustration. This chemical structure gives agarose the capacity to form gels that are very strong even at low concentrations. These gels have a macroreticular structure with a very open mesh which can be adjusted simply by varying the concentration of agarose. The macroreticule of the agarose gel is formed by hydrogen bonds, which makes the gel thermo-reversible, thus it melts upon heating. The hysteresis ­ difference between gelling and melting temperature ­ is greater than for any other hydrocolloid. In addition, the absence of ionic groups makes the gel a neutral structure, thus there is no interaction with hydrophilic macromolecules which migrate through the gel mesh. The gel is an efficient sieve for these particles.

Applications

All applications for agarose take advantage of the special characteristics of the macroreticular gel. It is used as a sieve or support through which biological macromolecules such as proteins or nucleic acids can pass. Larger particles, such as viruses and subcellular fragments, are also able to move through the gel network. Immunodiffusion. In this technique, macromolecules migrate and are precipitated in the gel by molecular diffusion. Electrophoresis. Agarose is suitable for the widest range of electrophoresis procedures as well as in immunoelectrophoresis and electrofocusing. Driven by electrostatic fields, the macromolecules migrate through the macroreticular structure. Gel Chromatography, Affinity Chromatography and Ion Exchange Chromatography. In these applications, the movement of macromolecules is caused by the displacement of solvent along the gel formed in microspheres. Supports for Biocatalysis. Agarose is derivatized and activated by organic synthesis to make supports for molecules with enzymatic activity. The capacity of gel beads as enzymatic support is much greater because enzymes can also be attached inside the beads. The structure is sufficiently open to allow the movement of coenzymes and substrates inside the gel. Solid Culture Media. Solid or semi-solid media are used to grow plant cells and tissues. Culture media prepared with agarose (instead of agar) can be used for strict autotrophic bacteria. Growth of Protein Crystals. The agarose gel regulates the diffusion of the protein molecules, allowing the formation of crystals suitable for crystallographic study.

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Agarose (electrophoresis grade)

NZYTech electrophoresis grade agarose is a standard type of agarose suitable fo the daily needs of DNA electrophoretic analysis

1800 bp

Features

Extraordinary mechanical resistance for reliable and easier handling Possibility of varying pore size in accordance with particle size by modifying the gel concentration Easy dilution in an appropriate buffer by standard boiling or microwaving Greater thermal stability due to high hysteresis (difference between gelling and melting temperatures) Excelent transparency of the gel and high visibility Exceptional low absorption of staining agents Absence of toxicity (polyacrylamide is toxic)

1000 bp 800 bp 600 bp 400 bp 200 bp

700 bp 600 bp 500 bp 400 bp 300 bp 200 bp

1x TBE buffer, 1%

23130 bp

2322 bp

Applications

High electrophoresis mobility Nucleic acid analytical and preparative electrophoresis Blotting assays Protein electrophoresis such as radial immunodifusion

1230 bp

517 bp 154 bp

1x TAE buffer, 1%

Specifications

Moisture Ash EEO (electroendosmosis) Sulphate Clarity 1.5% Gel Strength 1% (g cm-2) Gel Strength 1.5% (g cm-2) Gelling Temperature 1.5% (°C) DNase/RNase activity DNA resolution 1000 bp DNase/RNase Gel Background 7% 4% 0.05 ­ 0.13 0.1% 3% 1200 2500 36 ± 1.5 88 ± 1.5 None detected Finely resolved Very low

Cat No. MB02702 MB02703 MB02704

Pack Size 100 g 500 g 1000 g

Price 51,00 199,00 385,00

Purchase Online at: www.nzytech.com/site/Agaroses/Agarose-electrophoresis-grade

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Agarose (ultrapure grade)

NZYTech ultrapure grade agarose is a standard gelling/melting temperature agarose with high gel strength. This agarose is genetic quality tested which ensures that preparative electrophoresis can be performed and DNA recovered without damaging its properties and structure

Features

Extraordinary mechanical resistance for reliable and easier handling Possibility of varying pore size in accordance with particle size by modifying the gel concentration Easy dilution in an appropriate buffer by standard boiling or microwaving Greater thermal stability due to high hysteresis (difference between gelling and melting temperatures) Excelent transparency of the gel Low absorption of staining agents Absence of toxicity

1800 bp 1000 bp 800 bp 600 bp 400 bp 200 bp

700 bp 600 bp 500 bp 400 bp 300 bp 200 bp

1x TBE buffer, 1%

23130 bp

Applications

Analytical and preparative gel electrophoresis for nucleic acids Blotting assays Recovery of DNA fragments for further applications (enzymatic processing or cloning)

2322 bp 1230 bp

517 bp

154 bp

1x TAE buffer, 1%

Specifications

Cat No. MB05201 MB05202 Pack Size 100 g 500 g Price 88,00 299,00

Moisture Ash EEO (electroendosmosis) Sulphate Clarity 1.5% Gel Strength 1% (g cm-2) Gel Strength 1.5% (g cm-2) Gelling Temperature 1.5% (°C) DNase/RNase activity 7% 4% 0.05 ­ 0.13 0.1% 3% 1200 2500 36 ± 1.5 88 ± 1.5

Purchase Online at: www.nzytech.com/site/Agaroses/Agarose-ultrapure-grade

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(pulseAgarose (pulse-field grade)

NZYTech high molecular mass grade agarose is a linear polymer with a very high molecular weight, giving gels structures unlike those of traditional agaroses. This characteristic, added to the very low sulphate content, produces a strong intercatenary interaction, yielding a gel with very high gel strength and higher exclusion limit

Features

Extremely high gel strength allowing for lower gel concentrations (0.3%), enabling it to be used not only with high molecular weight nucleic acids, including chromossomes, but also with large sized particules like viruses and ribossomes High electrophoretic mobility. DNA mobility is greater when compared with the electrophoresis grade agarose. Electrophoresis times are reduced depending upon buffer and agarose concentration Easy dilution in an appropriate buffer by standard boiling or microwaving Greater thermal stability due to high hysteresis (difference between gelling and melting temperature) Low absorption of staining agents Absence of toxicity

40 kb 20 kb 10 kb 12 kb 10 kb 6 kb

1x TAE buffer, 0.5%

Applications

Conventional electrophoresis: can be used in a wide range of concentrations Pulse Field Gel Electrophoresis: because of its higher exclusion limit Blotting assays Agarose beads preparation Cell and enzyme immobilization Specifications

Moisture Ash EEO (electroendosmosis) Sulphate Clarity 1.5% Gel Strength 1% (g cm-2) Gel Strength 1.5% (g cm-2) Gelling Temperature 1.5% (°C) DNase/RNase activity DNA resolution 1000 bp DNase/RNase Gel Background 7% 0.25% 0.12 0.12% 4% 1800 3200 36 ± 1.5 88 ± 1.5 None detected Finely resolved Very low

Cat No. MB12201

Pack Size 500 g

Price 225,00

Purchase Online at: www.nzytech.com/site/Agaroses/Agarose-pulse-field-grade

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(LMAgarose (LM-ultrapure grade)

This agarose is a low gelling/melting temperature with genetic quality tested. Possessing high resolution capacity, finely resolves nucleic acid fragments from 50 bp to 2000 bp, especially PCR products. Due to its low gelling/melting temperatures LM-ultrapure agarose is compatible with In Gel applications (enzymatic processing of nucleic acids directly in remelted agarose) thus it is not necessary to recover DNA from agarose gels. The low viscosity of this agarose makes it possible for gels of high concentrations, even 6%, to be prepared easily. At lower concentration (<2%) gels are fragile and difficult to handle, so special care must be taken when working. The best concentration range for easy handling is 3-6%.

1800 bp 1000 bp 600 bp 400 bp

Features

Fine resolution: DNA fragments 2000 bp DNase/RNase activity non detected DNA binding none detected In-gel enzymatic processing Very low gel background

700 bp 500 bp 400 bp 300 bp 200 bp

1x TBE buffer, 4%

Applications

Analytical and preparative gel electrophoresis of small DNA fragments In-Gel applications

1353 bp 872 bp 310 bp 118 bp 72 bp

622 bp

110 bp

1x TBE buffer, 2%

Specifications

Cat No. MB12301 MB12302 Pack Size 100 g 500 g Price 165,00 785,00

Moisture Ash EEO (electroendosmosis) Sulphate Clarity 1.5% Gel Strength 1% (g cm-2) Gel Strength 1.5% (g cm-2) Gelling Temperature 1.5% (°C) DNase/RNase activity 7% 0.45% 0.13 0.12% 6% 800 3200 35 65

Purchase Online at: www.nzytech.com/site/Agaroses/Agarose-LM-ultrapure-grade

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General Applications and Separation Range

In the following table, you can find a summary of NZYTech agaroses and their recommended applications:

Analytical separation 2000 bp Analytical separation 2000 bp

Agarose

Preparative electrophoresis

PFGE

DNA typing

Blotting

Fine resolution

In Gel applications

Electrophores is grade Ultrapure grade Pulse-field grade LM-ultrapure grade

NZYTech agaroses provide a suitable system to analyze DNA fragments over a wide range of molecular weigths

200 400 600 800 12000 14000 16000 18000 25000 35000

1000

2000

4000

6000

8000

10000

Electrophoresis & Ultrapure agaroses 0.8% 1% 1.2% 1.5%

0.6%

Pulse-field agarose 0.5% 0.8% 1% 1.8%

0.3%

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DNA Stains

Portfolio

DNA Staining

Ethidium bromide (EtBr) is probably the most well-known dye used for visualizing DNA. Because of its chemical structure, it intercalates into the DNA strands forming fluorescent complexes that can be visualized under UV light. But Ethidium bromide is a potential carcinogen and mutagenic agent, so it must be handled with very special care and usually requires specific and exclusive bench space in order not to contaminate the entire laboratory. Because of all the health risks involved while working with EtBr, several alternative DNA stains have been developed during the recent years. Those alternatives are much less dangerous to work with and show a very good staining performance. NZYTech is proud to announce a portfolio of three alternative nucleic acid stains: GreenSafe, GreenSafe Premium and GreenSafe Direct Load. Their sensitive is similar to EtBr and agarose gel electrophoresis can be performed in the exact same way as with EtBr.

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GreenSafe

GreenSafe is a nucleic acid stain that can be used as a safer alternative to the traditional ethidium bromide stain for detecting nucleic acids in agarose gels. GreenSafe emits green fluorescence when bound to DNA or RNA. Downstream applications such as cloning can benefit from the use of GreenSafe as UV exposure may be minimized.

Features

Used for detecting dsDNA and RNA Non-toxic, non-mutagenic and non-carcinogenic As sensitive as ethidium bromide Convenient ready-to-use Compatible with a wide variety of gel reading instruments No hazardous waste

Agarose gel (1%, w/v) stained with GreenSafe

Applications

Incorporate in the agarose gel during its preparation Stain the gel after the run (poststaining)

The thickness of gel should be <0.5 cm. Repeatedly melting of gels containing GreenSafe may result in a lower sensitivity. GreenSafe is non-carcinogenic but may irritate skin and eyes. Please wear gloves while handling the product.

Cat No. MB08801

Pack Size 1 ml

Price 85,50

Purchase Online at: www.nzytech.com/site/DNA-loading-and-staining/GreenSafe

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GreenSafe Premium

GreenSafe Premium is an improved nucleic acid stain that can be used as a safer alternative to the traditional ethidium bromide for detecting nucleic acids in agarose gels. Besides the positive effects on your health, this dye shows a very high sensitivity, even for small fragments, as well as a superior signal to noise ratio. Furthermore, this dye has a reduced migration rate.

Features

High sensitivity even for small fragments Superior signal to noise ratio Used for detecting dsDNA and RNA Non-toxic, non-mutagenic and non-carcinogenic Convenient ready-to-use Compatible with a wide variety of gel reading instruments No hazardous waste

Agarose gel (1%, w/v) stained by GreenSafe Premium

Applications

Incorporate in the agarose gel during its preparation Stain the gel after the run (poststaining)

1 ml of GreenSafe Premium is sufficient for 17-25 litres of agarose. The thickness of the gel should be <0.5 cm. GreenSafe Premium is non-carcinogenic but may irritate skin and eyes. Please wear gloves while handling the product.

Cat No. MB13201

Pack Size 1 ml

Price 85,50

Purchase Online at: www.nzytech.com/site/DNA-loading-and-staining/GreenSafe

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GreenSafe Direct Load

GreenSafe Direct Load is a DNA stain designed to be applied directly in the sample(s). It includes a loading dye that allows the sample(s) to be promptly run in an agarose gel without having to add another product. GreenSafe Direct Load is a very high sensitivity dye and has an excellent signal noise ratio. Like the other GreenSafe products, it is a safer alternative to the traditional ethidium bromide stain.

Features

Direct gel loading High sensitivity Superior signal to noise ratio Used for detecting dsDNA, ssDNA and RNA Non-toxic, non-mutagenic and non-carcinogenic Compatible with a wide variety of gel reading instruments No hazardous waste

DNA samples with GreenSafe Direct Load were loaded onto 1% (w/v) agarose gel

Applications

Direct gel load: just mix the sample(s) with this dye at a 1:8 to 1:10 dilution rate (no need to add any other dyes to the gel matrix and or the running buffers)

GreenSafe Direct Load is impenetrable to latex gloves and cell membranes. GreenSafe Direct Load is non-carcinogenic but may irritate skin and eyes. Please wear gloves while handling the product.

Cat No. MB13301

Pack Size 1 ml

Price 85,50

Purchase Online at: www.nzytech.com/site/DNA-loading-and-staining/GreenSafe

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Customer Service Team

+351 213643514

www.nzytech.com

NZYTech, Lda. Estrada do Paço do Lumiar, Edifício E ­ R/C, 1649-038 Lisboa, Portugal Tel: +351 213643514 Fax: +351 217151168 Email: [email protected]

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Microsoft PowerPoint - NZYTech Brochure_Agaroses and DNA Stains.pptx