Team:Illinois/Protocols

From 2008.igem.org

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check [[Team:Illinois/Research articles|research articles]]
check [[Team:Illinois/Research articles|research articles]]
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===possible helpful links:===
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http://cgsc.biology.yale.edu/
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Frozen Competent E.Coli Cells. Methods and protocol for freezing E.Coli Bacteria Competent Cells in aliquots for later use re-use. http://www.ciwemb.edu/labs/koshland/Protocols/BACTERIA/ecolicells.ht
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Opimization of recombitant E. coli protein expression in growth medium
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http://minds.wisconsin.edu/handle/1793/7956
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===Host strain influences on supercoiled plasmid DNA production in Escherichia coli: Implications for efficient design of large-scale processes.===
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Yau SY, Keshavarz-Moore E, Ward J.
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Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: +44‐2076792961; fax: +44‐2079163943.
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Summary:
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* Set out to investigate if E. coli genotype plays a significant role in host strain selection for optimal processing of plasmid DNA based on both quality and quantity of supercoiling
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* 17 E. coli commercial and non-commercial strains were selected and their available genetic backgrounds were researched in the open literature. Growth characteristics of all the strains were considered and made impartial by using a common medium and growth condition platform.
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* Can screen the number of strains which are likely to give good productivity early on, before any further optimisation and verification is performed, both for small and large plasmids.
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* Found that high specific growth rates of some strains did not affect the level of DNA supercoiling but did influence the total plasmid yield, potentially an important aspect in the design of fermentation strategy. Interestingly, a few host/plasmid combinations result in what appears to be runaway plasmid replication.
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<br>
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*Potentially helpful for deciding strains to use to amplify plasmids
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<nowiki>-Bobak</nowiki>
===GPCRs===
===GPCRs===

Revision as of 22:38, 28 May 2008

Home The Team The Project Parts Submitted to the Registry Modeling Research Articles Notebook Current Tasks


Contents

Current Tasks and Subgroup Status

Yeast Culture

[http://www.protocol-online.org/prot/Model_Organisms/Yeast/Yeast_Culture/ Longterm storage and streaking protocols]

[http://www.mrw.interscience.wiley.com.proxy2.library.uiuc.edu/emrw/9780471142720/cp/cpmb/article/mb1302/current/pdf Growth and manipulation of yeast]

[http://openwetware.org/wiki/Yeast_culture_media Yeast culture media]

[http://www.protocol-online.org/prot/Model_Organisms/Yeast/Yeast_Transformation/ Yeast transformation protocols]

Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual (1998 edition available in Biology library, newer editions currently checked out)

[http://openwetware.org/wiki/Yeast Yeast Info on OWW] Contains General, Protocols, Materials, and References Info - Alzar

Directed Evolution

Malaria

E. coli

check research articles

possible helpful links:

http://cgsc.biology.yale.edu/

Frozen Competent E.Coli Cells. Methods and protocol for freezing E.Coli Bacteria Competent Cells in aliquots for later use re-use. http://www.ciwemb.edu/labs/koshland/Protocols/BACTERIA/ecolicells.ht

Opimization of recombitant E. coli protein expression in growth medium

http://minds.wisconsin.edu/handle/1793/7956

Host strain influences on supercoiled plasmid DNA production in Escherichia coli: Implications for efficient design of large-scale processes.

Yau SY, Keshavarz-Moore E, Ward J.

Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: +44‐2076792961; fax: +44‐2079163943.

Summary:

  • Set out to investigate if E. coli genotype plays a significant role in host strain selection for optimal processing of plasmid DNA based on both quality and quantity of supercoiling
  • 17 E. coli commercial and non-commercial strains were selected and their available genetic backgrounds were researched in the open literature. Growth characteristics of all the strains were considered and made impartial by using a common medium and growth condition platform.
  • Can screen the number of strains which are likely to give good productivity early on, before any further optimisation and verification is performed, both for small and large plasmids.
  • Found that high specific growth rates of some strains did not affect the level of DNA supercoiling but did influence the total plasmid yield, potentially an important aspect in the design of fermentation strategy. Interestingly, a few host/plasmid combinations result in what appears to be runaway plasmid replication.


  • Potentially helpful for deciding strains to use to amplify plasmids

-Bobak

GPCRs

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