Team:Paris/3 August 2009

From 2009.igem.org

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===Brain work===
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Meeting this morning for the lab work week. Discuss about the schedule algorithm for the iPhone application.  
Meeting this morning for the lab work week. Discuss about the schedule algorithm for the iPhone application.  
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<div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">
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<div class="caroline">
DID dying for membrane fluorescence microscopy
DID dying for membrane fluorescence microscopy
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<div class="experience">
*wash cell with MgSO4 then dying and incubation 30 min 25°C ->observation : 1/100 fluorescent cell and some vesicles ?!  
*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 ->observation : no significant (1/100)  
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->try another method : FM 4-64
->try another method : FM 4-64
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<div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">
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<div class="guillaume">
Miniprep digestion by XbaI and PstI
Miniprep digestion by XbaI and PstI
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*For [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P1]] pSB2K3, [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P2]] BBa_J61002, [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P3]] BBa_J32015, [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P4]] BBa_B0014, [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P5]] BBa_I14032, [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P7]] BBa_R0040, [[https://2009.igem.org/Team:Paris/Freezer#Plasmids:_MiniPrep_Products P8]] BBa_I0500.
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<div class="experience">
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*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
*Digestion Mix
**50µl Miniprep
**50µl Miniprep
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**1µl Loading (LD)
**1µl Loading (LD)
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Gel Migration
Gel Migration
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<div class="experience">
*[P1|P2|P3|P4|P5|Ladder 100bp|P7|P8]
*[P1|P2|P3|P4|P5|Ladder 100bp|P7|P8]
**P1 = pSB2K3 = plasmid backbone = 4399bp (expected weight)
**P1 = pSB2K3 = plasmid backbone = 4399bp (expected weight)
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<center>[[Image:100bp.png]][[Image:Digestion_030809.png]]</center>
<center>[[Image:100bp.png]][[Image:Digestion_030809.png]]</center>
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<div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">
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<div class="guillaume">
Culture ON for Miniprep
Culture ON for Miniprep
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</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px">
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*[S7] DH5&alpha;(g3p)  
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<div class="experience">
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*[https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S7 S7] DH5&alpha;(g3p)  
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==To do list==
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===To do list===
{|
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|miniprep digestion using Xba/Pst enzymes
|miniprep digestion using Xba/Pst enzymes
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{{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 →

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