CulGenex™

Synthetic Defined Media

Cat. no. G410 Synthetic Complete Agar, 15x100mm Plate, 26ml 10 plates/bag
Cat. no. G425 Synthetic Complete Agar minus Adenine and Histidine, 15x100mm Plate, 26ml 10 plates/bag

INTENDED USE

Hardy Diagnostics CulGenex™ Synthetic Defined Media are recommended for use as selective growth media for isolation of transformants and confirmation of auxotrophic phenotypes.

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

SUMMARY

The yeast Saccharomyces cerevisiae and Schizosaccharomyces pombe serve as model eukaryotic organisms for studying numerous cell processes. While minimal medium, also known as Synthetic Dextrose Minimal Medium, can support the growth of these yeasts; many strains routinely used in molecular biology contain well-characterized auxotrophic markers that cannot be grown on this medium. Auxotrophs lack the ability to produce essential components for the synthesis of nucleic acids or proteins. Identification and confirmation of auxotrophic strains are accomplished by replica plating on Synthetic Complete Agar and a medium lacking that nutritional requirement (typically an amino acid or nucleotide) referred to as "drop-out" media. Furthermore, many commonly used yeast vectors, including integrative vectors and episomal plasmids, contain markers for the synthesis of seletive amino acids and nucleotides. When auxotrophs are transformed with these vectors, the ability to grow in the absence of these molecules is restored.

Hardy Diagnostics CulGenex™ Synthetic Complete Agar contains the complete drop-out mixture of essential amino acids and nucleotides. Synthetic Complete Agar minus Adenine and Histidine drop-out plates contain all of the drop-out ingredients, minus the specified amino acids.

FORMULA

Ingredients per liter of deionized water:*

Synthetic Complete Agar:
Glucose 20.0gm
Yeast Nitrogen Base without Amino Acids 6.7gm
Leucine 10.0gm
Alanine 2.0gm
Arginine 2.0gm
Asparagine 2.0gm
Aspartic acid 2.0gm
Cysteine 2.0gm
Glutamine 2.0gm
Glutamic acid 2.0gm
Glycine 2.0gm
Histidine 2.0gm
Inositol 2.0gm
Isoleucine 2.0gm
Lysine 2.0gm
Methionine 2.0gm
Phenylalanine 2.0gm
Proline 2.0gm
Serine 2.0gm
Threonine 2.0gm
Tryptophan 2.0gm
Tyrosine 2.0gm
Uracil 2.0gm
Valine 2.0gm
Adenine 0.5gm
para-Aminobenzoic acid 0.2gm
Agar 20.0gm

In addition, Synthetic Complete Agar minus Adenine and Histidine do not contain these ingredients.

Final pH 5.6 +/- 0.2 at 25ºC.

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

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.


PRECAUTIONS


PROCEDURE

Please refer to appropriate references for recommended test proceduces. (1,2,5,6)

INTERPRETATION OF RESULTS

Interpretation of results will be indicated by growth, and variation in colony size dependent upon drop-out mixture used.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, applicator sticks, other culture media, incinerators, and incubators, etc., as well as serological and biochemical reagents, 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
Saccharomyces cerevisiae
ATCC ® 9763
A 24-72hr 35°C Aerobic Growth

USER QUALITY CONTROL


Physical Appearance

CulGenex™ Synthetic Defined Media should appear translucent, and light 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. Current Protocols in Molecular Biology . John Wiley and Sons, Inc. Malden, MA.

2. Bullock, G.R. and Petrusz, P. Techniques in Immunocytochemistry, Volumes 1, 2, 3 and 4, Academic Press, London.

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

4. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.

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

6. Walker, J.M. 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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