CulGenex™

Terrific Broth with Glycerol Powder

Cat. no. C8150 CulGenex™ Terrific Broth with Glycerol, 2L 80gm
Cat. no. C8151 CulGenex™ Terrific Broth with Glycerol 500gm
Cat. no. C8152 CulGenex™ Terrific Broth with Glycerol 2kg
Cat. no. C8153 CulGenex™ Terrific Broth with Glycerol 10kg
Cat. no. C8159 CulGenex™ Terrific Broth with Glycerol, 0.5L 6 pouches/pack

INTENDED USE

Hardy Diagnostics CulGenex™ Terrific Broth with Glycerol is recommended for use in cultivating recombinant strains of Escherichia coli .

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

Terrific Broth was developed by Tartoff and Hobbs in the attempt to increase the yield of plasmid DNA from transformed Escherichia coli strains. Terrific Broth has now become a popular choice in molecular biology laboratories as a medium for use in plasmid preparation. It is a highly enriched medium which produces higher yields of plasmids as a result of the increased concentration of peptone and yeast extract. Glycerol is also added to the formula as a carbohydrate source and unlike glucose, is not fermented to acetic acid. Peptone and yeast extract provide necessary nutrients and cofactors needed for the growth of recombinant strains of Escherichia coli . Yeast extract is added in a higher concentration to increase cell yields; and potassium phosphates are added to prevent cell death due to changes in pH.

FORMULA*

Gram weight per liter: 51.6gm/L
Yeast Extract 24.0gm
Tryptone 12.0gm
Dipotassium Phosphate 9.4gm
Glycerol 4.0gm
Monopotassium Phosphate 2.2gm

Final pH 7.2 +/- 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 media at 2-30ºC.


PRECAUTIONS


METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend 51.6gm of the dehydrated culture media in 1 liter of distilled or deionized water (25.8gm per 500ml).

2. Heat as necessary to dissolve completely.

3. Autoclave at 121ºC. for 15 minutes.

PROCEDURE

Refer to appropriate references for recommended test procedures. (1,2,5,6)

INTERPRETATION OF RESULTS

Growth is observed by turbidity and/or cloudiness of the broth medium.

LIMITATIONS


MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as autoclave, 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 24-48hr 35°C Aerobic Growth

User Quality Control


PHYSICAL APPEARANCE

CulGenex™ Terrific Broth with Glycerol powder should appear homogeneous, free-flowing, and very light to 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, and K. Struhl. 1994. Current Protocols in Molecular Biology . Greene Publishing Associates, Inc., Brooklyn, NY.

2. Lennox, E.S. 1955. Transduction of Linked Genetic Characters of the Host by Bacteriophage P1 . Virology; 1:190-206.

3. Luria, S.E., and J.W. Burrows. 1955. Hybridization Between Escherichia coli and Shigella . J. Bacteriol.; 74:461-476.

4. Luria, S.E., J.N. Adams, and R.C. Ting. 1960. Transduction of Lactose-Utilizing Ability Among Strains of E. coli and S. dysenteriae and the Properties of the Transducing Phage Particles . Virology; 12:348-390.

5. Miller, J.H. 1972. Experiments in Molecular Genetics . Cold Spring Harbor Laboratory. Cold Spring Harbor, New York.

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

7. Tartoff, K.D., and C.A. Hobbs. 1987. Improved Media for Growing Plasmid and Cosmid Clones. Bethseda Research Laboratories Focus; 9:12.


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

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