IGEM:IMPERIAL/2007/Calendar/2007-7-30

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We are in Week 4 of the iGEM project.


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Contents

Proposals

Task List

Students

Supervisors

Today's Schedule

Name Alex Anthony Ben Cheuk Dirk James Jerry Lucas Maira Peixuan
0900
0930
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Evening

Questions

Debriefing

The debrief session was attended by Professor Freemont. A number of issues were raised; these are described below.

Projects

The work done on the various projects was reported at the debriefing session.

Biofilm

  • General protocols available for E Coli experimentation were recorded.
  • Work was started on protocols specific to the Biofilm project
  • It was reported that 4 constructs would be employed. Also, it was suggested, that due to some parallels with last year's Imperial project, these established constructs could already be exploited.
  • More work needs to be done in terms of reporter mechanisms, e.g. DSRed.
  • This project proceeds by employing "test" constructs until the required HRP constructs are made available.

Problems/considerations

  • Can E Coli Polymerase be employed in the in-vitro/in-veso system, or is T7 the only possibility?
  • DSRed, was suggested as a possible reporter system; especially since this would already be characterized from one of the other projects. Are there other reasons which could/could not warrant its use in the Biofilm project? - especially, consider time-scale of operation.

HRP

  • The constructs to be pursued further were discussed.
  • Relevant protocols to be developed per device.

In-Veso

  • Email Christina from the "Vesicles" Division concerning considerations/issues/problems raised including:
    1. Stability/Rigidity of vesicles
    2. No. of ribosomes possibly accommodated by vesicles - in other words, size considerations.
    3. Polymerase restrictions - only T7 employable?
    4. Methods of fabrication
    5. Materials employed - particular w.r.t. phospholipid compostion/mixture used.
    6. Storage of vesicles, especially at temperatures of 4 degrees Celsius.
    7. Communication methods across membrane.
    8. Alternative means of forming pores within membrane, since a-hemolysin toxin is not permitted during our project work.
  • Also, included in this email, should be the relevant research papers on which our methods/project work to date has been based.

Cell-by-Date

  • Proposed new reporter mechanism, involving DSRed - inherent long degradation time.
  • Issues overlapping with In-Vitro/In-Veso systems, e.g. non-T7 polymerase, etc.
  • Cmv was initially proposed as a constitutive promoter; however, this can only be used successfully within mammalian cells; not prokaryotic cells.
  • The list of experiments to be conducted should be completed.
  • (other issues covered? input required)

Ambitions by end of week

  • Start production of media within lab.
  • Hopefully start cloning for E Coli System.
  • Organize division of labour between different projects, and, within the projects themselves, especially w.r.t labwork.
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