Neidhardt EZ Rich Defined

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==Background==
==Background==
This media is a slight variant on that defined by [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=4604283 Neidhardt ''et al'']. The modifications were made by [http://www.genome.wisc.edu/ Blattner ''et al''.]  This is a rich defined medium which should allow repeatable high growth rates.  The recipe from below is edited from the [http://www.genome.wisc.edu/resources/protocols/ezmedium.htm E. Coli Genome Project].  It would be our intention to use this as a standard media for all characterization work as a replacement for supplemented M9 media.
This media is a slight variant on that defined by [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=4604283 Neidhardt ''et al'']. The modifications were made by [http://www.genome.wisc.edu/ Blattner ''et al''.]  This is a rich defined medium which should allow repeatable high growth rates.  The recipe from below is edited from the [http://www.genome.wisc.edu/resources/protocols/ezmedium.htm E. Coli Genome Project].  It would be our intention to use this as a standard media for all characterization work as a replacement for supplemented M9 media.
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EZ Media has the following advantages -
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*Concentrations of the major nutrients can be varied independently (C, P, N, S)
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*Concentrations of all constituents are defined
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*Growth rate should be similar to that of 'E. coli' in rich media such as LB
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*Supports a high density of cell growth
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*Stable and gives reproducible results
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*Low background fluorescence, can be used for fluorescent measurements
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*Excellent pH buffering capacity
==Materials Sourcing & Usage==
==Materials Sourcing & Usage==

Revision as of 16:15, 4 January 2006

Contents

Background

This media is a slight variant on that defined by Neidhardt et al. The modifications were made by Blattner et al. This is a rich defined medium which should allow repeatable high growth rates. The recipe from below is edited from the E. Coli Genome Project. It would be our intention to use this as a standard media for all characterization work as a replacement for supplemented M9 media.

EZ Media has the following advantages -

  • Concentrations of the major nutrients can be varied independently (C, P, N, S)
  • Concentrations of all constituents are defined
  • Growth rate should be similar to that of 'E. coli' in rich media such as LB
  • Supports a high density of cell growth
  • Stable and gives reproducible results
  • Low background fluorescence, can be used for fluorescent measurements
  • Excellent pH buffering capacity

Materials Sourcing & Usage

  • We buy the two of the ingredients (10X ACGU and the 5X Supplement EZ) from Teknova because they are tedious to make. The Endy Lab has an account with Teknova (see Barry for details).
  • The kitchen makes up batches of 10X MOPS Mixture for us.
  • The kitchen also makes K2HPO4 for us.
  • Currently, because some of the ingredients are stored in our freezer we are making up the final media mix. In the future we may get the kitchen to make up the whole thing except for the carbon source so people can choose what they want to use. As listed in the protocol section, the carbon source should be glycerol to conform to the working SBWG media standards.
  • I have 4 L of the final media made up so people can test their experiments in this media before switching over entirely. Please see me if you want to get some --Bcanton 14:16, 7 Jun 2005 (EDT)
  • As the kitchen will be making the solutions on demand, please see me if you want more media made up.
  • Please feel free to share any feedback on this media via the discussion page or in person.

Protocol

  1. The following protocol makes 1 L of Media
  2. All of the ingredients have already been filter sterilized or autoclaved as appropriate.
  3. To a 1 L flask add each of the following ingredients listed in the table below.
  4. To conform to the working SBWG standardization, the 100x carbon source should be 40% Glycerol.
  5. Filter Sterilize with a .2μm filter.
1 10X MOPS Mixture 100 mL
2 0.132M K2HPO4(Dibasic) 10 mL
3 10X ACGU 100 mL
4 5X Supplement EZ 200 mL
5 Sterile H2O 580 mL
6 100x Carbon Source 10 mL
7 TOTAL 1 L

References

  • F. C. Neidhardt, P. L. Bloch, and D. F. Smith. 1974. Culture medium for enterobacteria. J Bacteriol 119(3): 736-747
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