AGAROSE PRODUCT INFORMATION
Introduction Agarose is a fraction extracted from agar-producing seaweeds and it is mainly responsible for the agaroses gelling power. It exhibits a high hysteresis (the difference between melting and gelling temperatures) making it an ideal media for separations techniques such as electrophoresis and chromatography within the fields of Molecular Biology and Biochemistry. Specifically, the gelling temperature ranges are 32 - 45°C and the melting temperature ranges are normally 80 - 95°C, although these can be modified when preparing products for specific uses. Agarose is a natural product that forms an inert matrix. Likewise, it is neutral and easily derivatizable, so it can easy be made to bind to proteins like enzymes, antigens or antibodies. The absence of toxicity makes working with agarose very safe.
Nowadays agarose is an essential tool in nucleic acid separation for Genetic Engineering, Cell Culture and Microbiology. The structure of the polysaccharide is a galactan, formed by linking agarobioses 1-3, 1-4, as shown in the illustration. This chemical structure gives agarose the capacity to form gels that are very resistant even at low concentrations. The macroreticule of the agarose gel is formed by hydrogen bonds, which makes the gel thermo-reversible, thus it melts after 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.
Properties § § § § § § § §
Dry powder, non hygroscopic Electrically neutral Biologically inert Can be stored for several years, if stored properly Free of toxicity (non neurotoxic as acrylamide) Can be autoclaved for sterile applications Easily derivatizable No free-radicals/additives needed for gelification
§ § § § § § § §
Macroporous matrix - allows molecular diffusion Easy preparation Thermoreversible, gels formed by hydrogen bonds Resistant, even at low concentrations Thermal stability Low absorption of staining agents-low background Pore size changes only by changing concentration Gels can be dried for recordkeeping
Applications § Inmunodiffusion: 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.
§ 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. § There are other scientific and technical applications that are continously appearing, like in cosmetics (additive, dermal filler, etc), for molds in a ceramic industry, etc.
Agarose Preparation AGAROSE DISSOLUTION 1. Dispersion: separation of the particles by the buffer preventing clumping. 2. Hydration: individual particles are surrounded by the solution (water, buffer); it is advisable to allow adiquate hydration time before heating for melting and dissolution. 3. Melting and dissolution: solid particles are re liquified. Different agarose types behave differently: There is no universal protocol for heating and dissolving agarose. Pore size is determined by concentration and agarose type used. An appropriate agarose and concentration should be chosen for each application . GEL PREPARATION: TIPS § Always use a beaker 2-4 times the volume of the solution. § Add agarose powder slowly into rapidly stirring buffer solution to avoid clumping. § The buffer solution should be cool for a good dispersion; if the buffer is warm, clumping is more likely. § Always allow agarose powder to hydrate in solution for a few minutes before heating - this allows a quicker and easier dissolution and reduces foaming. § Adjust time and power settings according to your microwave output strength. § Always wear appropriate protection: the microwaved solution can become overheated and foam when disturbed. § To prevent overheating: reduce microwave power, remove beaker after 1 min from the microwave and swirl it very gently and carefully. Place it back into the microwave and continue for the remaining minute or so. § For total agarose melting: boil the solution only long enough to acvhieve total dissolution. Check for “fish eyes” (incomplete dissolution). Overboiling can cause agarose hydrolysis and lower gel strength. § To avoid bubble formation: cool to 60ºC before pouring carefully into the gel cassette. § After pouring, allow the gel to cool gradually; rapid cooling will cause irregular gel matrix formation and band distortion during electrophoresis. § Low melt agarose gels need to stand for an additional 30 min or overnight at 4-8ºC to complete the gelling process. § Low melting or low percentage gels: it is important to run these gels in a cold buffer. High voltages can cause overheating of the buffer which can melt the gel. § Buffer composition can effect the gelling process, if agents that disrupt hydrogen bond formation are added to the buffer, melting temperature and gel strength will decrease, or even be completely inhibited. § Once the gel has set, saturate it with the buffer. The gel can be stored refrigerated for several days. § Agarose gels can be remelted and repoured several times without damage so that a large volume of agarose can be prepared and smaller portions taken from time to time.
AGAROSE MANUAL Hi Res Standard Agarose (AGD1,AGGQT,AGD1ME,AGD1HE) Hi Res Standard Agarose is available for different uses in 4 types: Low EEO*, M edium EEO, High EEO, and Hi-Res Standard GQT. GQT (Genetic Quality Tested) Agarose is similar to Hi-Res Standard I.e. a standard gelling /melting tem perature agarose with high gel strength, but also GQT (Genetic Quality Tested), certified to ensure that preparative electrophoresis can be performed and DNA recovered without dam aging its properties and structure. Hi-Res Standard GQT gels can be used in M olecular Biology techniques.
Features – Extraordinary mechanical resistance for more reliable and easier handling. – Possibility of varying pore size in accordance with particle size by modifying the gel concentration. – Easy preparation of the gel by simple dilution in aqueous buffers either by standard boiling or microwaving. – Greater thermal stability due to high hysteresis (difference between gelling and melting tem peratures). – Excellent transparency of the gel & visibility. – Exceptionally low absorption of staining agents. – Absence of toxicity (the alternative is polyacrylam ide which can be toxic).
A
B
Applications
C
23130 bp
1230 bp 2322 bp
517 bp
154 bp
1 2
1 2
1 2
Hi-Res Standard Agarose gels in IXTAE buffer A-0. 75% , B-1%, C1. 25% M arkers: lane 1 - Lam bda DNA. HindIII; lane 2 - pBR328DN A. BgII+pBR 328D NA. Hinfl. Electrophoresis conditions: subm arine gel, 2 hours, 4. 5 V/cm in 1 XTAE buffer.
Hi-Res Standard: with low EEO. High electrophoresis mobility. - Nucleic acid analytical and preparative electrophoresis. - Blotting. - Protein electrophoresis such as radial immunodiffusion. Hi-Res Standard GQT: with low EEO. - Analytical and preparative gel electrophoresis for nucleic acids ≥ 1000 bp. - Blotting assays. - Recovery of DNA fragments for further applications (enzymatic processing or cloning). Hi-Res Standard ME: with intermediate EEO. - Nucleic acids electrophoresis. - Protein electrophoresis (serum protein and immunoelectrophoresis). Hi-Res Standard HE: with high EEO. - Used in techniques such as serum protein, immunoelectrophoresis and counterimmunoelectrophoresis.
Specifications and Functional Tests Hi-Res Standard Moisture Ash EEO* Sulfate Clarity 1.5 % (NTU) Gel Strength 1% (g/cm 2 ) Gel Strength 1.5 % (g/cm 2 ) Gelling Tem perature 1. 5 % (°C) M elting Temperature 1. 5 % (°C) DNAse/ RNAse activity DNA resolution G 1000 bp Gel background *EEO (Electroendosmosis)
≤ 7% ≤ 0.4% 0. 05 - 0. 13 ≤ 0.1% ≤ 3 ≥ 1200 ≥ 2500 36 ± 1. 5 88 ± 1. 5 None detected Finely resolved Very low
Hi-Res Standard GQT ≤ 7% ≤ 0.4% 0. 05 - 0. 13 ≤ 0.1% ≤ 3 ≥ 1200 ≥ 2500 36 ± 1. 5 88 ± 1. 5 None detected Finely resolved Very low
Hi-Res Standard M E
Hi-Res Standard HE
≤ 7% ≤ 0.5% 0. 16 - 0. 19 ≤ 0.14% ≤ 4 ≥ 1000 ≥ 2200 36 ± 1. 5 88 ± 1. 5 None detected Finely resolved Very low
≤ 7% ≤ 1.0% 0. 23 - 0. 26 ≤ 0.2% ≤ 4 ≥ 750 ≥ 1200 36 ± 1. 5 88 ± 1. 5 None detected Finely resolved Very low
Hi-Gel Standard (AGD2)
Hi-Gel Standard Agarose has a higher gelling tem perature than Hi Res Standard Agarose and Low EEO*. This gives a higher thermal stability to the gels.
Features - Extraordinary mechanical resistance for more reliable and easier handling. - Possibility of varying pore size in accordance with particle size by modifying the gel concentration. - Easy preparation of the gel by simple dissolution in aqueous buffers either by standard boiling or microwaving. - Greater thermal stability due to high hysteresis (difference between gelling and melting tem peratures). - Excellent transparency of the gels. - Excellent elasticity and flexibility of the gels.
Specifications and Functional Tests
- Great capacity for derivatization and cross-linking, which allows coupling of enzymes, antigens
High Gel Standard
and other substances to the gel structure.
Moisture
- Exceptionally low absorption of staining agents. which can be toxic).
Applications Hi-Gel Standard: - Nucleic acid electrophoresis. - Protein electrophoresis (immunoelectrophoresis and counterelectrophoresis).
≤ 8%
Ash
- Absence of toxicity (the alternative is polyacrylamide
- Preparation of agarose beads.
D-2
≤ 0.4%
EEO* Sulfate Clarity 1.5% (NTU) Gel Strength 1% (g/cm 2 ) Gel Strength 1.5% (g/cm 2 ) Gelling Temperature 1. 5% (°C) M elting Tem perature 1. 5% (°C) DNAse/RNAse activity DNA resolution G1000bp Gel background
≤ 0.14 ≤ 0.2% ≤ 4 ≥ 900 ≥1200 42±1. 5 87±1. 5 None detected Finely resolved Very low
*EEO (electroendosmosis)
FP Rapid DNA (AGFP) Finger Printing DNA Agarose is a powerful tool in laboratories performing forensic testing, paternity determination, cell line verification, tissue typing, etc. FP DNA Agarose meets all requirements for DNA identity applications.
Features
Specifications and Functional Tests FP R a p i d DN A
- Low EEO. - High gel strength, forming easy–to-handle gels. - No DNA binding. - No DNAse and RNAse activity. - Clear and sharp bands. - High efficiency transfer for DNA (blotting). - No smearing. - No gel background. - No variability in agarose quality and performance
Moisture Ash Sulfate EEO* Gel Strength 1% (g/cm 2 ) Gelling Temperature 1. 5% (°C) M elting Tem perature 1. 5% (°C) DNAse/RNAse activity DNA binding DNA background DNA resolution
*EEO (electroendosmosis)
≤ 7% ≤ 0.4% ≤ 0.14% ≤ 0.13 ≥ 1400 36±15 88±1. 5 None detected None detected None detected Clean and sharp bands produced when a 23 kb DNA size Standard is electrophoresed transferred and probed.
Mega Base Rapid (D5)
Mega Base Rapid Agarose is a linear polymer with a very high molecular weight, giving gel structures unlike those of traditional agaroses.This characteristic, added to the very low sulfate content, produces an strong intercatenary interaction, yielding a gel with very high gel strength and higher exclusion limit.
Features - Extrem ely high gel strength allowing for lower gel concentrations (0. 3%), enabling it to be used not only with high molecular weight nucleic acids, including chromosomes, but also with large sized particles like viruses and ribosomes.
As we can see in the following photographs, Hi Res Standard Agarose is suitable for a wide variety of ranges, just by modifying its concentration.
- High electrophoretic mobility. DNA mobility is greater when com pared with D-1LE. Electrophoresis times are reduced depending upon buffer and agarose concentration used.
A
B
C
- Easy preparation of the gel by simple dissolution in aqueous buffers either by standard boiling or microwaving. - Greater thermal stability due to high hysteresis (difference between gelling and melting tem peratures).
12 Kb 10 Kb
40 Kb 30 Kb
6 Kb
20 Kb 4 Kb
- Exceptionally low absorption of staining agents. - Absence of toxicity (the alternative is polyacrylam ide which can be toxic).
10 Kb
Applications - Conventional Electrophoresis: can be used in a wide range of concentrations. - Pulsed Field Gel Electrophoresis: because of its higher exclusion limit, larger molecules can be separated. - Blotting.
1 2
1 2
1 2
Mega Base Rapid Agarose gels in 1 X TAE. A-0. 3%, B-05%, C-08%. Markers: lane 1-5 kb, lane 2-1 kb ladder. Electrophoresis conditions: submarine gel, 16 hours,
- Agarose Beads preparation.
D
E
F
23130 pb
Specifications and Functional Tests Moisture Ash EEO* Sulfate Clarity 1.5% (NTU) Gel Strength 1% (g/cm 2 ) Gel Strength 1.5% (g/cm 2 ) Gelling Temperature 1. 5% (°C) M elting Tem perature 1. 5% (°C) DNAse/RNAse activity DNA resolution > 1000 bp Gel background *EEO (electroendosmosis)
Mega Base Rapid ≤ 7% ≤ 0.25% ≤ 0.12 ≤ 0.12% ≤ 4 ≥ 1800 ≥ 3200 36±1. 5 88±1. 5 None detected Finely resolved Very low
6557 pb 1766 pb
1033 pb 2027 pb 653 pb
298 pb
1 2
1 2
1 2
Mega Base Rapid Agarose gels in 1 X TAE. D-0. 5%, E-1% , F-1. 5% . M arkers: lane 1-Lam bda DNA. HindIII, lane-2-pBR328DNA. Bgll+pBR328DN A. Hindfl. Electrophoresis conditions: subm arine gel, 2 hours, 4. 5 V/cm. in 1XTAE buffer.
8 Kb
A
Low Melt Clone
B
622 bp
(AGLMSVE)
Low Melt Clone Agarose is a low melting tem perature agarose with the highest resolving capacity for DNA fragments smaller than 1000 bp, especially PCR products ranging from 200 to 800 bp.This agarose is GQT (Genetic Quality Tested) certified. This ensures that In-Gel applications can be performed in remelted agarose, avoiding difficult DNA extraction steps. Low Melt Clone Agarose is ideal for digestion by agarase enzymes, making it very easy to recover small DNA fragments suitable for cloning or enzymatic processing. Low Melt Clone Agarose can be used at high concentrations, forming gels with excellent clarity and a higher sieving capacity than standard melting agaroses. Due to their high gel strength, Low Melt Clone Agarose gels are very easy to handle, even at concentrations as low as 2%.
1353 bp
160 bp
872 bp
110 bp
310 bp
34 bp 118 bp 72 bp
1 2
Applications ‐ Electrophoresis of DNA fragments ≤1000 bp. - In-Gel enzymatic processing (digestion, ligation, PCR). - Preparative electrophoresis. - Analysis and recovery of sm all DNA fragments for further applications.
Functional Tests -
C
DNA resolution: bands appear sharp and finely resolved. DNAse/RNAse activity: none detected. DNA binding: none detected. In-Gel enzymatic processing: passes test. Enzymatic degradation by agarase: passes test.
1 2
1 2
L o w M e l t C l o n e Agarose gels in 1XTBE buffer A-2% , B-3% , C-4%. M arkers: lane 1 - pBR322DN A. Mspl; lane 2 - øX174DNA . HaeIII Electrophoresis conditions: subm arine gel, 2 hours 30 min. , 4. 5 V/cm in 1XTBE buffer.
Specifications Low Melt Clone Moisture Ash EEO* Sulfate Gel Strength 4% (g/cm 2 ) Gelling Temperature 4% (°C) Melting Temperature 4% (°C)
≤ 5% ≤ 0.3% ≤ 0.10 ≤ 0.12% ≥ 1000 ≤ 35 ≤ 65
* EEO (electroendosm osis)
Novagel GQT
(AGNOVA)
A
NovaGel is a new low gelling /melting tem perature Agarose GQT grade certified. This agarose, with high resolution capacity, finely resolves nucleic acid fragments from 50 bp to 1000 bp, especially PCR products. Due to its low gelling /melting tem peratures NovaGel GQT 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 NovaGel GQT agarose makes it possible for gels of high concentrations, even 6% , to be prepared easily. At low er 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%.
Applications - Analytical and preparative gel electrophoresis of small DNA fragments. - In-Gel applications. - Analysis and recovery of small DNA fragments for further applications.
Functional Tests ‐ Fine resolution: DNA fragments ≤ 1000 bp. - DNAse/RNAse activity: none detected. - DNA binding: none detected. - In-Gel enzymatic processing: passes test. - Enzymatic degradation by agarose: passes test. - Gel background: very low after Et. Br. staining.
B
C 622 pb
1353 pb 872 pb
160 pb 110 pb
310 pb 34 pb
118 pb 72 pb
1 2 Novagel GQT Agarose gels in 1X TBE buffer. A-2% , B-3% , C-4% . M arkers: lane 1-pBR322 DN A. M spI, lane-2-ØX174 DN A. HaeIII. Electrophoresis conditions: subm arine gel, 2 hours, 4. 5 V/cm. in 1X TBE buffer.
Specifications Moisture Ash EEO* Sulfate Clarity 4% (NTU) Gel Strength 4% (g/cm 2 ) Gelling Temperature 4% ( o C) Melting Temperature 4% ( o C) * EEO (electroendosm osis)
NOVAGEL GQT ≤ 7% ≤ 0.45% ≤ 0.13 ≤ 0.12% ≤ 6 ≥ 800 ≤ 35 ≤ 65
Hi-Res Low Melt & Hi-Res Low Melt GQT (AGLM2,LMGQT) Low M elting (Hi-Res Low Melt) Agaroses are derivatized by organic synthesis which generates methoxylate groups from the basic agarose structure. The m ain properties of these agaroses are their low m elting and gelling tem peratures when com pared with standard agaroses.The low melting temperature allows for the recovery of undamaged nucleic acids at temperature lower that its denaturing temperature. The low gelling temperature assures the agarose will be in a liquid state at a temperature range where In-Gel manipulations can be performed without prior extraction of the DNA from the gel slice. Hi-Res Low Melt (GQT) Agarose is a low melting temperature agarose with the highest resolving capacity for large DNA fragments, ≥1000 bp, including PCR products. This agarose is GQT (Genetic Quality Tested) certified. This ensures that In‐Gel applications can be performed in remelted agarose, avoiding difficult DNA extraction steps. Hi-Res Low Melt ( GQT) Agarose is ideal for digestion by agarase enzymes, which makes it very easy to recover large DNA fragments suitable for cloning or enzymatic processing.
Features - Lower gel strength than standard agaroses. Even so, gels can be handled easily. - Higher clarity (gel transparency) than gels of standard agaroses. - Great sieving capacity. Hi-Res Low Melt Agaroses are classified in three categories, depending on the degree of derivatization. Gelling / melting tem peratures and gel strength are the most important differences.
Applications ‐ Electrophoresis of DNA fragments ≥1000 bp. - In-Gel enzymatic processing (digestion, ligation, PCR). - Preparative electrophoresis. - Analysis and recovery of large DNA fragments for further applications.
A
B
C
Functional Tests - DNA resolution: bands appear sharp and finely resolved. - DNAse/RNAse activity: none detected. - Gel background: very low after Et. Br. staining. Only for Hi-Res Low Melt GQT: - DNA binding: none detected. - In-Gel enzymatic processing: passes test. - Enzymatic degradation by agarase: passes test.
Specifications Moisture Ash EEO* Sulfate Clarity 1.5% (NTU) Gel Strength 1.5% (g/cm 2 ) Gelling Tem perature 1. 5% (°C) Melting Temperature 1.5% (°C) DNAse/RNAse activity DNA resolution G1000 bp Gel background *EEO (electroendosmosis)
Hi-Res Low Melt ≤ 7% ≤ 0.4% ≤ 0.12 ≤ 0.12% ≤ 4 ≥ 500 24-28 ≤ 65.5 None detected Finely resolved Very low
12 kb
5 kb
1 kb
Hi-Res Low Melt GQT Agarose at different concentrations. A-0. 75% , B-1% and C-1. 25% . M arker: 1kb ladder, 0. 5 µ g/lane. Running conditions: 1XTAE buffer, 4, 5V/cm, 2 hours 30 min.
Hi-Res Super +
(AGMS4)
A molecular screening agarose for improved resolution of DNA fragm ents with 500 bp or less, especially sized–primer fragments. At 3 % concentration, H i - Re s S u p e r + Agarose gives a resolution of DNA fragm ents similar to gels made with polyacrylamide at concentrations of 8 %. W hile H i - Re s S u p e r + may be dissolved carefully by microwaving, gels are best prepared by autoclaving.
Features
1
- Excellent resolution of DNA fragm ents lower than 500 bp, specially sm aller sized–primer fragm ents. - Forms a very clear, transparent gel, even at concentrations of 5% or higher. - Efficient mechanical handling at all concentrations. The chances of gel breaking or cracking when handled are greatly minimized.
500 bp 320 bp 190 bp 124 bp
Functional Tests
100 bp
- DNA resolution: bands appear sharp and finely resolved. - DNAse/RNAse activity: none detected. - Gel background: very low after Et. Br. staining.
80 bp
60 bp
Specifications
40 bp
Hi-Res Super + Moisture Ash EEO* Sulfate Clarity (NTU) Gel Strength (g/cm 2 ) Gelling Temperature (°C) Melting Temperature (°C)
2 3
3%
5%
≤ 7% ≤ 0.3% ≤ 0.12 ≤ 0.11%
20 bp
≤ 6 ≥ 500 ≤ 31 ≤ 76
≥ 1000
H i Re s S u p e r + Agarose gel, 4% in 0. 5XTBE buffer. M arkers: lane 1- M olecular weigth marker VIII (Roche) lane 2-M olecular weigth marker V (Roche) lane 3-10 bp ladder. Electrophoresis conditions: submarine gel, 2 hours 30 min. , 4. 5 V/cm in 0. 5XTBE buffer.
*EEO (electroendosmosis)
MetaGel (AGMETAMS6) M e ta G e l Agarose is a high quality agarose specially formulated for molecular screening. W ith M e ta G e l Agarose we can bring you an agarose with an improved efficiency resolution of sm all DNA fragments and PCR products.
Features
1 2 3
- High resolution capacity close to the resolution of polyacrylam ide gels. - Improved clarity of the gel, enhancing visualization, even at high concentrations. - High gel strength which enables an easy handling even when used at lower concentrations.
Functional Tests -
DNA resolution: bands appear sharp and finely resolved. DNAse/RNAse activity: none detected. Gel background: very low after Et. Br. staining. DNA binding: very low.
Specifications
1000 bp 800 bp 600 bp 400 bp
200 bp
100 bp
MetaGel Moisture Ash EEO* Sulfate Clarity (3%) (NTU) Gel strength (3%) (g/cm 2 ) Gelling Temperature (3%) (°C) Melting Temperature (3%) (°C) *EEO (electroendosmosis)
≤ 7 % ≤ 0.3 % ≤ 0.12 ≤ 0.1 % ≤ 4 ≥ 800 ≤ 35 ≤ 75
25 bp
M e taGel 3% Agarose gel in 1XTAE buffer. M arkers: lane 1 – 25 bp ladder lane 2 – M olecular weight marker V. lane 3 - 100 bp ladder Electrophoresis conditions: submarine gel, 2h 30 min, 4. 5 V/cm in 1XTAE buffer.
Hi-Res Super (AGMS8) An agarose for molecular screening that improves resolution of small DNA fragments and PCR products. We have produced H i - Re s S u p e r Agarose for applications that require efficient separation of small DNA fragments and PCR products.
Features - High resolution of short PCR and DNA fragm ents. - Improved clarity of the gel, enhancing visibility. - Better handling than competitive products because of a stronger gel structure and higher gel strength. The chances of gels breaking or cracking when handled are greatly minimized, even with lower concentrations of agarose. - High gel strength allows use in blotting. 1000 bp
Functional Tests -
1 2 3 4
600 bp 400 bp
DNA resolution: bands appear sharp and finely resolved. DNAse/RNAse activity: none detected. Gel background: very low after Et. Br. staining. DNA binding: very low.
300 bp 200 bp
100 bp 80 bp
Specifications
60 bp
1.5 %
Hi-Res Super
Moisture Ash EEO* Sulfate Clarity (NTU) Gel Strength (g/cm 2 ) Gelling Temperature (°C) Melting Temperature (°C)
40 bp
3%
≤ 7% ≤ 0.35% ≤ 0.12 ≤ 0.11%
20 bp
≤ 5 ≥ 600
H i Re s S u p e r Agarose gel, 3% concentration in 1XTAE buffer. M arkers: lane 1- 250 bp ladder. lane 2-100 bp ladder. lane 3-m olecular weight marker V (Roche). lane 4-10 bp ladder. Electrophoresis conditions: subm arine gel, 2 hours, 4. 5 V/cm in 1XTAE buffer.
≥ 1500 ≤ 35.5 ≤ 80
* EEO (electroendosmosis)
Hi Strength Analytical
(AGMS12)
This molecular screening agarose is designed to have a larger gel network than M S-8 and is recommended for the separation of DNA fragm ents smaller than 1500 bp. Gels made with H i S t re n gt h A n a l y t i ca l have higher gel strength than competitive products. The gel is exceptionally firm but still flexible when handled, minimizing the danger of cracking or breaking. H i S t re n gt h A n a l y t i c a l has the sam e melting and gelling tem perature as regular agaroses, allowing faster and easier preparation of gels. H i S t re n gt h A n a l y t i ca l also gives excellent resolution at concentrations of F1 %. This agarose is recom mended for all analytical applications, especially when DNA is recovered for subsequent use in enzymatic procedures. 1 2 3 4
Functional Tests -
DNA resolution: bands appear sharp and finely resolved. DNAse/RNAse activity: none detected. Gel background: very low after Et. Br. staining. Blotting: very good transference for DNA fragm ents 154 – 2176 bp in 4 % gels. DNA binding: very low.
2176 bp 1500 bp 1000 bp
500 bp
Specifications Hi Strength Analytical
Moisture Ash EEO* Sulfate Clarity (NTU) Gel Strength (g/cm 2 ) Gelling Temperature (°C) Melting Temperature (°C) * EEO (electroendosmosis)
1.0%
1.5 %
4.0%
≤ 7% ≤ 0.35% ≤ 0.12 ≤ 0.11%
267 bp
124 bp
≤ 5 ≥ 2000
≥ 4200 ≤ 40.5 ≤ 93
H i S t re n gt h A n a l y t i c a l Agarose gel, 2% concentration in 0. 5XTBE buffer. M arkers: Lane 1- pBR328DNA. Bgll+pBR328DNA. Hinfl. Lane 2 - 100 bp ladder. Lane 3 - pBR322DNA. M spl. Lane 4 - pBR322DNA. Haelll. Electrophoresis conditions: subm arine gel, 2 hours, 4. 5
AGTC Bioproducts Ltd (Trading as National Diagnostics) Unit 4 Fleet Buisness Park, Itlings Lane, Hessle, East Riding of Yorkshire, HU13 9LX Phone: 01482646020 Fax:01482646013 E-mail: Office@agtcbioproducts.com www.agtcbioproducts.com