CulGenex™

2 x YT Broth

Cat. no. C9160 CulGenex™ 2xYT Broth 62gm
Cat. no. C9161 CulGenex™ 2xYT Broth 500gm
Cat. no. C9162 CulGenex™  2xYT Broth 2kg
Cat. no. C9163 CulGenex™  2xYT Broth 10kg

INTENDED USE

Hardy Diagnostics CulGenex™ 2xYT Broth is recommended for use as a general purpose growth medium for the cultivation of recombinant strains of Escherichia coli and the propagation of M13 bacteriophage.

SUMMARY

2xYT Agar (2x Yeast Extract Tryptone Agar) was developed for the growth and maintenance of recombinant strains of Escherichia coli used for genetic research. (1,3,5,6) The medium is also ideal for the propagation of M13 bacteriophage for sequencing and antibody phage display research. (3-5)

Antibody-based immunologic research is useful for identifying, isolating or eliminating cells with particular characteristics related to various diseases, and phage display is a useful technique for antibody selection for this purpose. (4) From these technologies, large and diverse libraries of recombinant antibodies can be generated, typically for molecules associated with pathological conditions, and screening by antibody phage display may lead to new and more potent drug discoveries. (2,4)

Components of 2xYT Agar provide nitrogen and growth factors that allow bacteriophage to reproduce in large quantities, without exhausting resources from the host cell. E. coli also grows well in this medium due to readily available amino acids, nucleotide precursors, vitamins and other metabolites present for cell growth. (1,5)

FORMULA*

Gram weight per liter: 31 gm/L
Tryptone 16.0gm
Yeast Extract 10.0gm
Sodium Chloride 5.0gm

Final pH 6.8 +/- 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 beige.

Store the prepared culture media at 2-8ºC.


PRECAUTIONS


METHOD OF PREPARATION FOR DEHYDRATED PRODUCT(S)

1. Suspend 31gm of the dehydrated culture medium in one liter of distilled or deionized water.

2. Heat to boiling and mix to dissolve completely.

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

4. Allow media to cool to 50ºC before adding antibiotics or additional supplements.

PROCEDURE and INTERPRETATION OF RESULTS

For information on procedures and interpretation of results, consult listed references. (1-6)

LIMITATIONS


MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as autoclaves, 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 ® 23724
A 24-48 hrs 35°C Aerobic Growth

User Quality Control


Physical Appearance

CulGenex™ 2XYT Broth powder should appear homogeneous, free-flowing, and light beige in color. The prepared medium should appear translucent, slightly opalescent, 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. Bullock, G.R. and Petrusz, P. 1989. Techniques in Immunocytochemistry, Volumes 1, 2, 3 and 4, Academic Press, London.m

3. 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.

4. O'Brien, P.M and R. Aitken. 2002. Antibody Phage Display . The Humana Press, Inc. Clifton, NJ.

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

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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