CulGenex™

AGAROSE POWDER

Cat. no. C8740 CulGenex™ Agarose LE, Molecular Biology Grade 100gm
Cat. no. C8741 CulGenex™ Agarose LE, Molecular Biology Grade 500gm
Cat. no. C8750 CulGenex™ Agarose, Fine Resolution, PCR Grade 25gm
Cat. no. C8751 CulGenex™ Agarose, Fine Resolution, PCR Grade 125gm
Cat. no. C8760 CulGenex™ Agarose, Low Melt 25gm
Cat. no. C8761 CulGenex™ Agarose, Low Melt 125gm

INTENDED USE

Hardy Diagnostics CulGenex™ Agarose LE, Molecular Biology Grade is recommended for use in the preparation of an electrophoresis gel for the analysis of nucleic acids and proteins.

Hardy Diagnostics CulGenex™ Agarose, Fine Resolution, PCR Grade is recommended for use in the preparation of an electrophoresis gel intended to improve the resolution of small DNA fragments and PCR products.

Hardy Diagnostics CulGenex™ Agarose, Low Melt is recommended for use in the preparation of an electrophoresis gel with a lower melting and gelling temperature.

SUMMARY

Agarose is a neutral polysaccharide derived from agar, which is prepared from species of red marine algae, Gellidium and Gracilariae . Agar may be separated, using an acetylation process, into two different polysaccharides: agarose and aparopectin. (1) This results in the formation of an acidic polymer (agaropectin) and a neutral polymer (agarose). (2)

Agarose is not soluble in cold water, but will dissolve in boiling water or buffer. It is an ideal gel matrix for diffusion and fractionation of particles as small as one nucleotide. (3) The finished gel is an anti-convection medium, which is biologically inert and well suited for electrophoresis of nucleic acids, lipoproteins, lactic dehydrogenase isoenzymes, serum proteins, glycoproteins, heparin, acid mucopoplysaccharides, bacterial proteins, and some viruses. (4) Other uses include chromatography and immunological applications such as agarose gel diffusion, radial immunodiffusion, immunoelectrophoresis, electroimmunoassay, and counterelectrophoresis.

Agarose LE, Molecular Biology Grade has low electroendosmosis (EEO) and high mobility. The gelling temperature of this product is 34.5 to 37.5ºC. while the melting temperature is 86.5 to 89.5ºC.

Agarose, Fine Resolution, PCR Grade has a modified chemical structure of the polymer in order to improve the clarity, enhance differentiation of short fragments, and increase the gel strength, even in low agarose concentrations (especially useful for blotting). The gelling temperature of the Fine Resolution Agarose at a concentration of 3% is approximately 35.5ºC. while the melting temperature is 80ºC.

Agarose, Low Melt has a lower melting (65.5ºC. at 1.5%) and gelling temperature (24-28ºC. at 1.5%). Specifically, for separating DNA fragments greater than or equal to 1,000 base pairs. The methoxylate groups generated from organic synthesis of the basic agarose structure. allows for an increased clarity and decreased gel strength. This Agarose may also be used for tissue culture, cell culture and viral plaque assays.

There is no detectable amount of DNase or RNase in all CulGenex™ Agarose products.

STORAGE AND SHELF LIFE

Store the sealed container(s) containing dehydrated culture medium at 2-30ºC. Dehydrated medium is very hygroscopic. Keep container tightly sealed. Protect dehydrated media from moisture and light. The dehydrated media should be discarded if it is not free-flowing or if the color has changed from its original white.


PRECAUTIONS


METHOD OF PREPARATION

Refer to appropriate references for specific concentrations of agarose/ethidium bromide and formulas for TAE/TBE buffers.

1. In a flask, add Agarose to distilled water or a suitable buffer (TBE or TAE). For example, a 1% agarose gel with a total volume of 50mL would contain 0.5gm of Agarose.

2. Heat to boiling to dissolve completely. This may be done in a microwave and evaporated water might need to be replaced.

3. In order to later visualize DNA fragments through UV fluorescence, ethidium bromide may be added to the dissolved agarose.

4. Cool and pour into electrophoresis cast. Allow sufficient time for solidification before use.

LIMITATIONS

In order to increase resolution of small fragments with the Fine Resolution Agarose (PCR Grade) gels should be stored at 4-8ºC. for 30 minutes prior to use.


MATERIALS REQUIRED BUT NOT PROVIDED

Standard supplies and equipment such as flasks, microwaves etc., are not provided.

USER QUALITY CONTROL


PHYSICAL APPEARANCE

CulGenex™ Agarose powder should appear homogeneous, free-flowing, and white in color.

REFERENCES

1. Araki, C. 1937. Acetylation of Agar-like Substances of Gelidium amansii . L. J. Chem. Soc. Japan ; 58:1338-1350.

2. Barteling, S.J. 1969 A Simple Method for the Preparation of Agarose. Clin. Chem. ; 15:1002-1005.

3. Griess, G. et al. 1993. The Relationship of Agarose Gel Structure to the Sieving of Spheres during Agarose Gel Electrophoresis. Biophysical Journal. ; Vol. 65:138-148.

4. Santos, G. 1990. A Manual for the Processing of Agar from Gracilaria . No. 5. Regional Small-Scale Coastal Fisheries Development Project. Manila, Philippines.


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