CulGenex™

SUPER BROTH POWDER

Cat. no. C1519 CulGenex™ Super Broth Powder, 0.5L 6 pouches/pack

INTENDED USE

Hardy Diagnostics CulGenex™ Super Broth Powder is recommended for use in cultivating high densities of recombinant strains of Escherichia coli to increase the yield of plasmid DNA.

This dehydrated culture medium is a raw material intended to be used in the making of prepared media products, which will require further processing, additional ingredients, or supplements.

SUMMARY

Super Broth is a nutrient rich medium used in molecular genetics research to increase yields during plasmid or protein production. The medium was developed by Douglas Hanahan in 1983 and is a modification of the commonly used LB Broth medium intended to improve the production of plasmid DNA from E. coli strains. (3)

Bacterial transformation requires the formation of competent cells, which permit the introduction of foreign DNA into cells. Competent cells need a nutrient rich, isotonic environment in order to survive transformation. (3,5) Super Broth is formulated to support the growth of competent cells: the medium contains elevated levels of tryptone and yeast extract, making it rich in peptides, amino acids, vitamins and cofactors, which allow transformed cells to recover and grow; sodium chloride provides essential ions. Consequently, Super Broth supports the growth of logarithmic phase cells and results in higher cell densities versus traditional LB Broth media. (3-6) As an additional carbon source, glycerol may be added to the medium since, unlike glucose, glycerol is not fermented to acetic acid during cell growth.

Hardy Diagnostics CulGenex™ Super Broth Powder is packaged in single use pouches ready to yield 500ml of a final broth solution upon the addition of water. The product is available as a package of six pouches.

FORMULA*

Gram weight per liter: 57.0gm/L
Tryptone 32.0gm
Yeast Extract 20.0gm
Sodium Chloride 5.0gm

Final pH 7.0 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

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

Store the prepared culture media at 2-8ºC.


PRECAUTIONS


METHOD OF PREPARATION FOR DEHYDRATED PRODUCT(S)

Note: If a solid medium is desired, 15.0gm/L of agar may be added to the medium. Glycerol (5ml/L) may be added to the medium as an additional carbon and energy source. For optimal binding of lambda phage, 10ml of a 20% maltose solution (0.2% maltose) may be added to the medium to induce the lambda receptor (LamB) on host cells.

1. Suspend one pouch (0.5L) of the dehydrated culture media in 500ml of distilled or deionized water. Stir to mix thoroughly.

2. Heat as necessary to dissolve completely.

3. Dispense into desired autoclavable containers. If preparing agar, dispense after autoclaving.

4. Sterilize in the autoclave at 121ºC. for 15 minutes.

5. Cool to 45-50ºC.

PROCEDURE AND INTERPRETATION OF RESULTS

For information on procedures and interpretation of results, consult listed references.

LIMITATIONS

Maltose induction of the lambda receptor is not recommended when making liquid lysate phage stocks because liberated phage will bind to cellular membrane fragments containing a high concentration of phage receptors.


MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as flasks, petri dishes, autoclaves, scales, incinerators, and incubators, etc., are not provided.

QUALITY CONTROL

Hardy Diagnostics tests each lot of commercially manufactured media using appropriate quality control microorganisms and quality specifications as outlined on the Certificates of Analysis (CofA).

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Escherichia coli
ATCC ® 25922
A 24hr 35°C Aerobic Growth; turbidity

User Quality Control


Physical Appearance

CulGenex™ Super Broth Powder should appear homogeneous, free-flowing, and light tan in color. The prepared media should appear clear, and light to medium amber in color.

REFERENCES

1. Ausubel, F.M., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, K. Struhl, Editors. 2010. Current Protocols in Molecular Biology . John Wiley and Sons, Inc. Malden, MA.

2. Cseke, L.J., P.B. Kaufman, G.K. Podila, and C.J. Tsai. 2004. Handbook of Molecular and Cellular Methods in Biology and Medicine . CRC Press. Taylor & Francis LLC. Boca Raton, FL.

3. Hanahan, D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Bio .; 166(4):557-80.

4. Sambrook and Russell. 2001. Molecular Cloning: A Laboratory Manual , 3rd ed. Cold Spring Harbor Laboratory Press. Woodbury, New York.

5. Tartof, K.D. and C.A. Hobbs. 1987. Improved Media for Growing Plasmid and Cosmid Clones. Bethesda Research Laboratories Focus .; 9:12.

6. Walker, J.M. 1984. Methods in Molecular Biology . The Humana Press Inc. Clifton, NJ.


ATCC is a registered trademark of the American Type Culture Collection.

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