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

We are in Week 4 of the iGEM project.

<calendar> name=IGEM:IMPERIAL/2007/Calendar date = 2007/08/01 view=threemonths format=%name/%year-%month-%day weekstart=7 </calendar>

Today's Schedule

Name	Alex	Anthony	Ben	Cheuk	Dirk	James	Jerry	Lucas	Maira	Peixuan
0900
0930
1000
1030
1100
1130
1200
1230
1300
1330
1400
1430
1500
1530
1600
1630
1700
1730
1800
Evening

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.