CRITERION™ LURIA BROTH BASE, MILLER

Cat. no. C6320 CRITERION™ Luria Broth Base, Miller 31gm
Cat. no. C6321 CRITERION™ Luria Broth Base, Miller 500gm
Cat. no. C6322 CRITERION™ Luria Broth Base, Miller 2kg
Cat. no. C6323 CRITERION™ Luria Broth Base, Miller 10kg
Cat. no. C6324 CRITERION™ Luria Broth Base, Miller 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Luria Broth Base, Miller is used for maintaining and propagating Escherichia coli in molecular microbiology procedures with or without added glucose.

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

CRITERION™ Luria Broth Base, Miller is based on the Luria Broth formula described by Miller for the growth and maintenance of Escherichia coli strains used in molecular biology procedures.

Luria Broth Base is a nutritionally rich medium designed for growth of pure cultures of recombinant strains. Escherichia coli is grown of pure cultures on LB Medium. Some plasmid vectors may replicate to high copy numbers without selective amplification. Some vectors do not replicate so freely, and need to be selectively amplified. Chloramphenicol can be added to inhibit host synthesis and as a result, prevent replication of the bacterial chromosome.(2)

Luria Broth Base, Miller contains one tenth and one twentieth the sodium chloride level of the Lennox and Miller formulations of LB Agar.(1-3) This allows the researcher to select the optimal salt concentration for a specific strain. The medium may be aseptically supplemented with glucose if desired.

Peptides and peptones are provided by pancreatic digest of casein. Vitamins, including B vitamins, and certain trace elements are provided by yeast extract. Sodium ions for transport and osmotic balance are provided by sodium chloride.

FORMULA

Gram weight per liter: 15.5gm/L
Peptic Digest of Casein 10.0gm
Yeast Extract 5.0gm
Sodium Chloride 0.5gm

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 bottle(s) containing 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 CULTURE MEDIA

1. Suspend 15.5gm of the dehydrated culture media in 1 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. Cool to 45-50ºC. and aseptically add 10ml sterile 20% glucose solution and mix thoroughly.

PROCEDURE AND INTERPRETATION OF RESULTS

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

LIMITATIONS

MATERIALS REQUIRED BUT NOT PROVIDED

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

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Escherichia coli
ATCC ® 25922
A 18-24hr 35°C Aerobic Growth

User Quality Control

PHYSICAL APPEARANCE

CRITERION™ Luria Broth Base, Miller powder should appear homogeneous, free-flowing, and light tan in color. The prepared media should appear clear, opalescent, and light to very light amber in color.

REFERENCES

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

2. Sambrook, J., E.F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

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


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

060616gr