Team:Paris/3 August 2009

From 2009.igem.org

(Difference between revisions)
(→To do list)
(→Lab work)
 
(27 intermediate revisions not shown)
Line 4: Line 4:
==NoteBook==
==NoteBook==
-
{| style="color:#1b2c8a;background-color:#0;" cellpadding="0" cellspacing="1" border="0" bordercolor="#fff" width="100%" align="left"
+
{{Paris2009_Calendar}}
-
!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=06 }}
+
{{Paris2009_Calendar_Link|2_August_2009|4_August_2009}}
-
!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=07 }}
+
<center> '''August 3rd''' </center>
-
!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=08 }}
+
-
!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=09 }}
+
-
!align="center"|{{ #calendar: title=Team:Paris |year=2009 | month=10 }}
+
-
|}
+
<html>
<html>
Line 19: Line 15:
</html>
</html>
-
 
+
===Brain work===
-
==Brain work==
+
Meeting this morning for the lab work week. Discuss about the schedule algorithm for the iPhone application.
-
 
+
-
 
+
-
edit please ^^
+
Line 33: Line 26:
</html>
</html>
-
 
+
===Lab work===
-
==Lab work==
+
<div class="caroline">
-
 
+
DID dying for membrane fluorescence microscopy
-
edit please ^^
+
</div>
-
 
+
<div class="experience">
-
 
+
*wash cell with MgSO4 then dying and incubation 30 min 25°C ->observation : 1/100 fluorescent cell and some vesicles ?!
 +
*dying in medium, incubation 30min 37°C ->observation : no significant (1/100)
 +
*dying in medium, incubation 30min 37°C, wash with MgSO4 ->observation : no significant (1/100)
 +
->try another method : FM 4-64
 +
</div>
 +
<div class="guillaume">
 +
Miniprep digestion by XbaI and PstI
 +
</div>
 +
<div class="experience">
 +
*For [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1] pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P2 P2] BBa_J61002, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P3 P3] BBa_J32015, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P4 P4] BBa_B0014, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P5 P5] BBa_I14032, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P7 P7] BBa_R0040, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P8 P8] BBa_I0500.
 +
*Digestion Mix
 +
**50µl Miniprep
 +
**2µl XbaI
 +
**2µl PstI
 +
**0,6µl BSA
 +
**6µl Buffer n°2
 +
*2h at 37°C
 +
*Verification by migration on 1% Agarose gel with BET
 +
**5µl of digestion solution
 +
**1µl Loading (LD)
 +
</div>
 +
<div class="guillaume">
 +
Gel Migration
 +
</div>
 +
<div class="experience">
 +
*[P1|P2|P3|P4|P5|Ladder 100bp|P7|P8]
 +
**P1 = pSB2K3 = plasmid backbone = 4399bp (expected weight)
 +
**P2 = BBa_J61002 = plasmid backbone = (will not be digest by this enzyme)
 +
**P3 = BBa_J32015 = pelB = 66bp
 +
**P4 = BBa_B0014 = Double terminater = 95bp
 +
**P5 = BBa_I14032 = pLac = 37bp
 +
**P7 = BBa_R0040 = tetR repressible promoter = 54bp (weak but present, cf other photos)
 +
**P8 = BBa_I0500 = pBad/araC = 1210bp
 +
<center>[[Image:100bp.png]][[Image:Digestion_030809.png]]</center>
 +
</div>
 +
<div class="guillaume">
 +
Culture ON for Miniprep
 +
</div>
 +
<div class="experience">
 +
*[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S7 S7] DH5&alpha;(g3p)
 +
</div>
<html>
<html>
</div>
</div>
Line 46: Line 79:
</html>
</html>
 +
===To do list===
-
==To do list==
+
{|
-
 
+
|- style="background: #ccccff; text-align: center;"
-
''Work for the day and the week :''
+
|width=100px| Matricule
-
 
+
|width=800px| TODO
-
 
+
|- style="background: #59eb26;text-align: center;"
-
*Vicard : planification of all the steps necessary for the implementation of the ClyA device in our strains (today) ; search for articles on fusion of vesicules (with Pierre, Luc and Guillaume).
+
|Luc
-
 
+
|SNARE Construction
-
 
+
|- style="background: #efe715;text-align: center;"
-
*Soufiane : design of oligos -> FecR, FecA, SNARE?, sequencing oligo. First days in the lab.
+
|Romain
-
 
+
|Design oligo Fec A
-
 
+
|- style="background: #d8279c;text-align: center;"
-
*Pierre & ChristopheC. : modelling -> Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system  
+
|Charlotte
-
 
+
|design of oligos: FecR, FecA, SNARE?, sequencing oligo
-
 
+
|- style="background: #fbfbfb;text-align: center;"
-
*Caroline : coloration of cells ; protocols for the lab -> control of the emission of vesicules....
+
|Stoff
-
 
+
|Dev schedule algorithm
-
 
+
|- style="background: #0824f5;text-align: center;"
-
*Sylvain : minipreps. Check if the use of starch is possible ; problem : find a system to help the starch enter into the periplasm, use of LacY -> noise.
+
|Chris
-
+
|modelling: Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system  
-
 
+
|- style="background: #06ff00;text-align: center;"
 +
|Lisa
 +
|Oligo Tol/Pal / Oligo Sequencing / Tol/Pal Protocol verification / Keio / Theme A writting
 +
|- style="background: #b40ecd;text-align: center;"
 +
|Caroline
 +
|coloration of cells / protocols for the lab: control of the emission of vesicules....
 +
|- style="background: #ff9966;text-align: center;"
 +
|Souf
 +
|design of oligos: FecR, FecA, SNARE?, sequencing oligo
 +
|- style="background: #79f4f8;text-align: center;"
 +
|Vicard
 +
|planification of all the steps necessary for the implementation of the ClyA device in our strains / search for articles on fusion of vesicules
 +
|- style="background: #e35050;text-align: center;"
 +
|Pierre
 +
|modelling: Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system
 +
|- style="background: #bababa;text-align: center;"
 +
|Sylvain
 +
|minipreps. Check if the use of starch is possible ; problem : find a system to help the starch enter into the periplasm, use of LacY -> noise
 +
|- style="background: #009900;text-align: center;"
 +
|Guillaume
 +
|miniprep digestion using Xba/Pst enzymes
 +
|}
-
Use of autotransporters as an altenative to the snare device.
+
{{Paris2009_Calendar_Link|2_August_2009|4_August_2009}}

Latest revision as of 13:27, 10 August 2009

Contents

NoteBook

June
MTWTFSS
[http://2009.igem.org/Team:Paris/1_June_2009 1] [http://2009.igem.org/wiki/index.php?title=Team:Paris/2_June_2009&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Paris/3_June_2009&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Paris/4_June_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Paris/5_June_2009&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Team:Paris/6_June_2009&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Paris/7_June_2009&action=edit 7]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/8_June_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Paris/9_June_2009&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Paris/10_June_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Paris/11_June_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Paris/12_June_2009&action=edit 12] [http://2009.igem.org/Team:Paris/13_June_2009 13] [http://2009.igem.org/wiki/index.php?title=Team:Paris/14_June_2009&action=edit 14]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/15_June_2009&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Team:Paris/16_June_2009&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Team:Paris/17_June_2009&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Team:Paris/18_June_2009&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Paris/19_June_2009&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Paris/20_June_2009&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Paris/21_June_2009&action=edit 21]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/22_June_2009&action=edit 22] [http://2009.igem.org/Team:Paris/23_June_2009 23] [http://2009.igem.org/wiki/index.php?title=Team:Paris/24_June_2009&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Paris/25_June_2009&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Team:Paris/26_June_2009&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Paris/27_June_2009&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Paris/28_June_2009&action=edit 28]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/29_June_2009&action=edit 29] [http://2009.igem.org/Team:Paris/30_June_2009 30]
July
MTWTFSS
    [http://2009.igem.org/wiki/index.php?title=Team:Paris/1_July_2009&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Paris/2_July_2009&action=edit 2] [http://2009.igem.org/Team:Paris/3_July_2009 3] [http://2009.igem.org/wiki/index.php?title=Team:Paris/4_July_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Paris/5_July_2009&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=Team:Paris/6_July_2009&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Paris/7_July_2009&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Team:Paris/8_July_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Paris/9_July_2009&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Paris/10_July_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Paris/11_July_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Paris/12_July_2009&action=edit 12]
[http://2009.igem.org/Team:Paris/13_July_2009 13] [http://2009.igem.org/Team:Paris/14_July_2009 14] [http://2009.igem.org/Team:Paris/15_July_2009 15] [http://2009.igem.org/Team:Paris/16_July_2009 16] [http://2009.igem.org/Team:Paris/17_July_2009 17] [http://2009.igem.org/Team:Paris/18_July_2009 18] [http://2009.igem.org/Team:Paris/19_July_2009 19]
[http://2009.igem.org/Team:Paris/20_July_2009 20] [http://2009.igem.org/Team:Paris/21_July_2009 21] [http://2009.igem.org/Team:Paris/22_July_2009 22] [http://2009.igem.org/Team:Paris/23_July_2009 23] [http://2009.igem.org/Team:Paris/24_July_2009 24] [http://2009.igem.org/Team:Paris/25_July_2009 25] [http://2009.igem.org/Team:Paris/26_July_2009 26]
[http://2009.igem.org/Team:Paris/27_July_2009 27] [http://2009.igem.org/Team:Paris/28_July_2009 28] [http://2009.igem.org/Team:Paris/29_July_2009 29] [http://2009.igem.org/Team:Paris/30_July_2009 30] [http://2009.igem.org/Team:Paris/31_July_2009 31]
August
MTWTFSS
          [http://2009.igem.org/Team:Paris/1_August_2009 1] [http://2009.igem.org/Team:Paris/2_August_2009 2]
[http://2009.igem.org/Team:Paris/3_August_2009 3] [http://2009.igem.org/Team:Paris/4_August_2009 4] [http://2009.igem.org/Team:Paris/5_August_2009 5] [http://2009.igem.org/Team:Paris/6_August_2009 6] [http://2009.igem.org/Team:Paris/7_August_2009 7] [http://2009.igem.org/Team:Paris/8_August_2009 8] [http://2009.igem.org/Team:Paris/9_August_2009 9]
[http://2009.igem.org/Team:Paris/10_August_2009 10] [http://2009.igem.org/Team:Paris/11_August_2009 11] [http://2009.igem.org/Team:Paris/12_August_2009 12] [http://2009.igem.org/Team:Paris/13_August_2009 13] [http://2009.igem.org/Team:Paris/14_August_2009 14] [http://2009.igem.org/Team:Paris/15_August_2009 15] [http://2009.igem.org/Team:Paris/16_August_2009 16]
[http://2009.igem.org/Team:Paris/17_August_2009 17] [http://2009.igem.org/Team:Paris/18_August_2009 18] [http://2009.igem.org/Team:Paris/19_August_2009 19] [http://2009.igem.org/Team:Paris/20_August_2009 20] [http://2009.igem.org/Team:Paris/21_August_2009 21] [http://2009.igem.org/Team:Paris/22_August_2009 22] [http://2009.igem.org/Team:Paris/23_August_2009 23]
[http://2009.igem.org/Team:Paris/24_August_2009 24] [http://2009.igem.org/Team:Paris/25_August_2009 25] [http://2009.igem.org/Team:Paris/26_August_2009 26] [http://2009.igem.org/Team:Paris/27_August_2009 27] [http://2009.igem.org/Team:Paris/28_August_2009 28] [http://2009.igem.org/Team:Paris/29_August_2009 29] [http://2009.igem.org/Team:Paris/30_August_2009 30]
[http://2009.igem.org/Team:Paris/31_August_2009 31]
September
MTWTFSS
  [http://2009.igem.org/Team:Paris/1_September_2009 1] [http://2009.igem.org/Team:Paris/2_September_2009 2] [http://2009.igem.org/Team:Paris/3_September_2009 3] [http://2009.igem.org/Team:Paris/4_September_2009 4] [http://2009.igem.org/Team:Paris/5_September_2009 5] [http://2009.igem.org/Team:Paris/6_September_2009 6]
[http://2009.igem.org/Team:Paris/7_September_2009 7] [http://2009.igem.org/Team:Paris/8_September_2009 8] [http://2009.igem.org/Team:Paris/9_September_2009 9] [http://2009.igem.org/Team:Paris/10_September_2009 10] [http://2009.igem.org/Team:Paris/11_September_2009 11] [http://2009.igem.org/Team:Paris/12_September_2009 12] [http://2009.igem.org/Team:Paris/13_September_2009 13]
[http://2009.igem.org/Team:Paris/14_September_2009 14] [http://2009.igem.org/Team:Paris/15_September_2009 15] [http://2009.igem.org/Team:Paris/16_September_2009 16] [http://2009.igem.org/Team:Paris/17_September_2009 17] [http://2009.igem.org/Team:Paris/18_September_2009 18] [http://2009.igem.org/Team:Paris/19_September_2009 19] [http://2009.igem.org/Team:Paris/20_September_2009 20]
[http://2009.igem.org/Team:Paris/21_September_2009 21] [http://2009.igem.org/Team:Paris/22_September_2009 22] [http://2009.igem.org/Team:Paris/23_September_2009 23] [http://2009.igem.org/Team:Paris/24_September_2009 24] [http://2009.igem.org/Team:Paris/25_September_2009 25] [http://2009.igem.org/Team:Paris/26_September_2009 26] [http://2009.igem.org/Team:Paris/27_September_2009 27]
[http://2009.igem.org/Team:Paris/28_September_2009 28] [http://2009.igem.org/Team:Paris/29_September_2009 29] [http://2009.igem.org/Team:Paris/30_September_2009 30]
October
MTWTFSS
      [http://2009.igem.org/Team:Paris/1_October_2009 1] [http://2009.igem.org/Team:Paris/2_October_2009 2] [http://2009.igem.org/Team:Paris/3_October_2009 3] [http://2009.igem.org/Team:Paris/4_October_2009 4]
[http://2009.igem.org/Team:Paris/5_October_2009 5] [http://2009.igem.org/Team:Paris/6_October_2009 6] [http://2009.igem.org/Team:Paris/7_October_2009 7] [http://2009.igem.org/Team:Paris/8_October_2009 8] [http://2009.igem.org/Team:Paris/9_October_2009 9] [http://2009.igem.org/Team:Paris/10_October_2009 10] [http://2009.igem.org/Team:Paris/11_October_2009 11]
[http://2009.igem.org/Team:Paris/12_October_2009 12] [http://2009.igem.org/Team:Paris/13_October_2009 13] [http://2009.igem.org/Team:Paris/14_October_2009 14] [http://2009.igem.org/Team:Paris/15_October_2009 15] [http://2009.igem.org/Team:Paris/16_October_2009 16] [http://2009.igem.org/Team:Paris/17_October_2009 17] [http://2009.igem.org/Team:Paris/18_October_2009 18]
[http://2009.igem.org/Team:Paris/19_October_2009 19] [http://2009.igem.org/Team:Paris/20_October_2009 20] [http://2009.igem.org/Team:Paris/21_October_2009 21] [http://2009.igem.org/Team:Paris/22_October_2009 22] [http://2009.igem.org/Team:Paris/23_October_2009 23] [http://2009.igem.org/Team:Paris/24_October_2009 24] [http://2009.igem.org/Team:Paris/25_October_2009 25]
[http://2009.igem.org/Team:Paris/26_October_2009 26] [http://2009.igem.org/Team:Paris/27_October_2009 27] [http://2009.igem.org/Team:Paris/28_October_2009 28] [http://2009.igem.org/Team:Paris/29_October_2009 29] [http://2009.igem.org/Team:Paris/30_October_2009 30] [http://2009.igem.org/Team:Paris/31_October_2009 31]

← Yesterday - Tomorrow →

August 3rd

Brain work

Meeting this morning for the lab work week. Discuss about the schedule algorithm for the iPhone application.


Lab work

DID dying for membrane fluorescence microscopy

  • wash cell with MgSO4 then dying and incubation 30 min 25°C ->observation : 1/100 fluorescent cell and some vesicles ?!
  • dying in medium, incubation 30min 37°C ->observation : no significant (1/100)
  • dying in medium, incubation 30min 37°C, wash with MgSO4 ->observation : no significant (1/100)

->try another method : FM 4-64

Miniprep digestion by XbaI and PstI

  • For P1 pSB2K3, P2 BBa_J61002, P3 BBa_J32015, P4 BBa_B0014, P5 BBa_I14032, P7 BBa_R0040, P8 BBa_I0500.
  • Digestion Mix
    • 50µl Miniprep
    • 2µl XbaI
    • 2µl PstI
    • 0,6µl BSA
    • 6µl Buffer n°2
  • 2h at 37°C
  • Verification by migration on 1% Agarose gel with BET
    • 5µl of digestion solution
    • 1µl Loading (LD)

Gel Migration

  • [P1|P2|P3|P4|P5|Ladder 100bp|P7|P8]
    • P1 = pSB2K3 = plasmid backbone = 4399bp (expected weight)
    • P2 = BBa_J61002 = plasmid backbone = (will not be digest by this enzyme)
    • P3 = BBa_J32015 = pelB = 66bp
    • P4 = BBa_B0014 = Double terminater = 95bp
    • P5 = BBa_I14032 = pLac = 37bp
    • P7 = BBa_R0040 = tetR repressible promoter = 54bp (weak but present, cf other photos)
    • P8 = BBa_I0500 = pBad/araC = 1210bp
100bp.pngDigestion 030809.png

Culture ON for Miniprep

  • S7 DH5α(g3p)

To do list

Matricule TODO
Luc SNARE Construction
Romain Design oligo Fec A
Charlotte design of oligos: FecR, FecA, SNARE?, sequencing oligo
Stoff Dev schedule algorithm
Chris modelling: Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system
Lisa Oligo Tol/Pal / Oligo Sequencing / Tol/Pal Protocol verification / Keio / Theme A writting
Caroline coloration of cells / protocols for the lab: control of the emission of vesicules....
Souf design of oligos: FecR, FecA, SNARE?, sequencing oligo
Vicard planification of all the steps necessary for the implementation of the ClyA device in our strains / search for articles on fusion of vesicules
Pierre modelling: Tol-Pal system, delay system, first equations in biocham ; use of mathlab. Think about the transduction system
Sylvain minipreps. Check if the use of starch is possible ; problem : find a system to help the starch enter into the periplasm, use of LacY -> noise
Guillaume miniprep digestion using Xba/Pst enzymes


← Yesterday - Tomorrow →

Retrieved from "http://2009.igem.org/Team:Paris/3_August_2009"