Melbourne/meeting 27 September 2007

From 2007.igem.org

< Melbourne(Difference between revisions)
 
Line 30: Line 30:
*Phil: Gas vesicles progress/design/plan - needs?
*Phil: Gas vesicles progress/design/plan - needs?
-
**Ligation P48J (SpeI & PstI digested) to PCR(GvpL) underway
+
**Ligation P48J (SpeI & PstI digested) to PCR(GvpL) -Failed
-
**Ligation P48J (SpeI & PstI digested) to PCR(GvpB to GvpU inclusive)(XbaI and PstI digested) underway
+
**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
**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)
**Other parts required from kit->glycerol stocks: B034 (P1-3O) , B0015 (P1-1I or P3-3O), J61034 (P4-8F or 8H)
Line 43: Line 54:
**Suggested Chris contact Lisa Hayes in chem eng re promotion.
**Suggested Chris contact Lisa Hayes in chem eng re promotion.
-
*Phil: Administrative Timetable: (4 Weeks to go)
+
*Phil: Administrative Timetable: (3 Weeks to go)
*Chris: Next years Comp Plan/preparations  
*Chris: Next years Comp Plan/preparations  
Line 57: Line 68:
==5:00 End ==
==5:00 End ==
 +
 +
==Igem Administrative Timetable:==
 +
 +
{|border="1"
 +
 +
|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.
 +
 +
 +
 +
{|border="1"
 +
 +
!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,
 +
 +
|}

Latest revision as of 04:29, 27 September 2007

Contents

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.


IndividualDesired 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,
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