From 2007.igem.org

Contents 1 3:30 Action points from last meeting: 2 3:35 Status Reports: (5 minutes each including any discussion) 3 3.55 New Agenda Items: 4 5:00 End 5 Igem Administrative Timetable: 6 Documentation wiki status 115+ hours work outstanding:

3:30 Action points from last meeting:

• Alisa: Get plan for blue light on wiki (from 10 Aug). Alisa has done a plan but this is not on Wiki.

3:35 Status Reports: (5 minutes each including any discussion)

• Pat : Red light receptor progress/design/plan- needs?
  • M30109: Must construct from scratch plan to be produced by Pat.
    • craig seqenced Cph8 from M30109 first part present
    • Cph8 from initial plate not functional
  • This is then drives RBS-ComP-term
    • RBS-ComP -done
  • Also required is ConstitutiveProm-RBS-ComA-term
    • RBS-ComA-term -done

• Jan & Alisa: Blue light receptor design/plan progress - needs?
  • ComP and SopII conbined at variouse points by using 4 primer stiching:
    • Comp P forward
    • SopII reverse
    • ComP-SoPII overlap sense
    • ComP-SopII overlap antisense
  • Note that there are 7 different stiching pairs that result in combination at different points.-Done
  • The product is then combined with RBS -Done
  • Sequencing being performed Friday
  • Then transcription terminator
  • The constitutive promotor to create protein generator part.
  • Also Required is reporter construct PsfPromoter-RBS-LacZ-term
  • Only three colonies pSFR8 grew in LB

• Phil: Gas vesicles progress/design/plan - needs?
  • Ligation P48J (SpeI & PstI digested) to PCR(GvpL) -Failed
  • Ligation P48J (SpeI & PstI digested) to PCR(GvpB to GvpU inclusive)(XbaI and PstI digested)-Failed
    • high background vector only control, and low ligation colony counts (Repeated unfortunately).
      • Think single cut of vector being repaired in cell for background.
      • Think Ligation producing linear that cant recircularise reducing background in ligation plates.
      • Think circular ligation of large plasmid rare and not affecting results significantly.
      • Ran test on P48J digestion 1uL 1 hour 37deg showed PstI incomplete in buffer 2 + BSA
      • Performed P48J 16hr 37deg double digest & Sequencial digest in buffer 2 then 3 plating to check background, with and without quick ligase.
    • Smaller GvpL plasmid showed *10 colony numbers cf background with overnight ligation protocol.
    • No increase in numbers with 5 minute ligation.
      • Think ligase affecting transformation efficiency.
      • Running test of effect of ligase on transformation efficiency of control plasmid pUC18.

• • Buoyancy experiment needs repeating
  • Other parts required from kit->glycerol stocks: B034 (P1-3O) , B0015 (P1-1I or P3-3O), J61034 (P4-8F or 8H)
  • Pat to provide these.

3.55 New Agenda Items:

• Sally Grass (chemical and biochemical engineering):
  • interested in signal transduction.
  • moving to lab in bio21.
  • will help promote next years comp
  • Suggested Chris contact Lisa Hayes in chem eng re promotion.

• Phil: Administrative Timetable: (3 Weeks to go)

• Chris: Next years Comp Plan/preparations
  • Slide prepared

• Alisa: To make competent cells to replace Joes

• Alisa: Tshirt should be simple based on Melb Uni logo, will propose a design

• Paul: suggested Ian Potter foundation may provide travel funding

• Next meeting: <b/> 27th September (Thurs) 3.30PM.

5:00 End

Igem Administrative Timetable:

12 October	Final team rosters due. Jamboree attendance fees due $100USD per team member USD$225 for others.
19 October	Registration with late fee USD $50 goes off line.
26 October	Project and wiki & part documentation frozen, DNA received USA.
3-4 November	iGEM Competition Jamboree, MIT, USA.

.

• re prism :Edmund optics:VIS60 Schott veril linear variable interference filter SGD$1600 25mm wide 60mm long linear 400nm-700nm (contact sheet)

N47-279 eqilateral prism 35mm long 35mm face SGD$162

Documentation wiki status 115+ hours work outstanding:

• • Priority should be on plans and experimental report analysis and conclusions-----Alisa .

• • Equipement:

• • • MissingLas3000 method (in lab notebook) 2h

• • • Missing DNA measurement protocol (eppendorf) 1h

• • Reagents pages incomplete: 24h

• • • Some location pictures not finished or uploaded. 12h

• • • Some newer reagents not entered. 8h

• • Secondary reagents pages not made:

• • • LB, Plates, TAE , NZY+ Broth 3h

• • Protocol part:

• • • No connections from Protocols to reagents. 3h

• • • Ligation protocol missing 2h

• • Lab notebook: no entries after 19 JUly 16h

• • • Format uses too many lines to see connections between days. 2h

• • • Latter part missing links 2h

• • • Can table of contents be restricted to date level? 4h

• • Experimental reports:

• • • Lacks discussion/conclusion 10h

• • • Ligation experiment missing

• • • Lattest few gells not marked up and entered. 20h

• • Project overview link broken 1h

• • Background:

• • • Blue light theory missing , 5h

• • • Red photo sensor to GFP test harness missing. 5h

• • Plan: Only Gas vesicles plan entered.

Individual	Desired time committment
Pat	About 4h/wk Monday afternoon or Tuesday morning
Alisa	About 4h/wk - Thursday afternoon
Jan	About 10h/pw - flexible weekday mornings
Phil	About 18h/pw - Thursday morning, Friday, Saturday or Sunday,