Team:Paris/6 August 2009

From 2009.igem.org

(Difference between revisions)
(→Lab work)
(→Molecular Biology)
 
(39 intermediate revisions not shown)
Line 15: Line 15:
</html>
</html>
-
==Brain work==
+
===Lab work===
 +
====Microscope====
 +
<div class="caroline">
 +
FM4-64 dying MG4 and Kayo delete Tol R
 +
</div>
 +
<div class="experience">
 +
protocol n°2 (05/08/09) but using PBS
 +
->nearly the same as MgSO4
-
edit please ^^
+
protocol n°2 (05/08/09)but 1ml of cells and 1µl of dye, one with MgSO4 and one with PBS
-
 
+
->not enough  dye
-
<html>
+
</div>
</div>
-
<div id="paris_content_boxtop">
+
====Molecular Biology====
 +
<div class="vicard">
 +
Gel Migration
</div>
</div>
-
<div id="paris_content">
+
<div class="experience">
-
</html>
+
*[Ladder 1kb|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A1 A1]|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A2 A2]|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3]]using 1% agarose during 30min
 +
**[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A1 A1] = Tg3p = 274bp (utiliser le ladder 100bp)
 +
**[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A2 A2] = TE3 = 1036bp
 +
**[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3] = ClyA (with RBS and poly G linker = 987bp
-
==Lab work==
 
-
<div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">
+
<center>[[Image:KR000634.JPG]][[Image:A3_cut.JPG]]</center>
-
FM4-64 dying MG4 and Kayo
+
-
</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px">
+
-
protocol n°2 (05/08/09) but using PBS
+
-
->nearly the same as MgSO4
 
-
protocol n°2 (05/08/09)but 1ml of cells and 1µl of dye, one with MgSO4 and one with PBS
+
*[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A1 A1] and [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A2 A2] didn't work we don't see it on the gel because it's not in E.Coli.
 +
*[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3] didn't migrate very well
 +
**[Ladder 1kb|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5]|nothing|[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5]|nothing] using 1% agarose during 30min
 +
**[https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5] = TolRII = 279bp
-
->not enough  dye
 
-
</div>
 
-
<div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">
+
<center>[[Image:KR000637.JPG]][[Image:A5_coupe.JPG]]</center>
-
Gel Migration
+
-
</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px">
+
-
*[Ladder 1kb|A1|nothing|A2|nothing|A3]using 1% agarose during 30min
+
-
**A1 = Tg3p = 274bp (utilisé le ladder 100bp)
+
-
**A2 = TE3 = 1036bp
+
-
***A1 and A2 didn't work
+
-
**A3 = ClyA (with RBS and poly G linker = 987bp
+
-
**[for more detail[https://2009.igem.org/Team:Paris/Freezer_PCR_products]]
+
-
<center>[[Image:KR000634.JPG]][[Image:A3coupe.JPG]]</center>
+
-
*[Ladder 1kb|nothing|A5|nothing|A5|nothing] using 1% agarose during 30min
+
-
**A5=TolRII=279bp[for more detail[https://2009.igem.org/Team:Paris/Freezer_PCR_products]]
+
-
<center>[[Image:KR000637.JPG]][[Image:A5coupé.JPG]]</center>
+
</div>
</div>
-
<div style="background-color:#7B96B3; border:1px solid #333333; font-weight:bold; padding:3px 5px; margin-bottom:1px">
+
<div class="vicard">
Purification and Digestion
Purification and Digestion
-
</div><div style="background-color:#E0E9EF; border:1px solid #333333; padding:3px 5px; margin-bottom:10px">
+
</div>
-
*Purification of A3, A4, A5, A6
+
<div class="experience">
 +
*Purification of [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A3 A3], [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A4 A4] directly in micro-column, [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A5 A5], [https://2009.igem.org/Team:Paris/Freezer_PCR_products#A6 A6] using gel then micro-column
*Digestion with XbaI and PSTI
*Digestion with XbaI and PSTI
</div>
</div>
-
Writing tomorow...
+
<div class="guillaume">
-
*Nouvelles Miniprep pour P1(pSB2K3), P13(pSB1A3), P14(BBa_K136050) => Nulles à refaire pousser ON et miniprep a refaire demain
+
New Miniprep
-
*ON pour S8(pSB2K3), S29(pSB1A3), S30(BBa_K136050)
+
</div>
-
*Digestion P1(pSB2K3), P2(BBa_J61002), P7(Bba_R0040) 2h37 par respectivement XP, XS, XP
+
<div class="experience">
-
*Migration sur gel pour verifier 1kb, P1, P2, P7, 100bp
+
*New Miniprep for [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1]: pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P13 P13]: pSB1A3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P14 P14]: BBa_K136050 (RBS tetR)
-
*Transformation 1L7 BBa_J24679 AmpR+ dans DH5a
+
*Result: Just good for trash. Make new ON culture.
 +
</div>
 +
<div class="guillaume">
 +
Digestion by XbaI/PstI (XP) or XbaI/SpeI (XS)
 +
</div>
 +
<div class="experience">
 +
*[https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1]: pSB2K3 by XP
 +
*[https://2009.igem.org/Team:Paris/Freezer_Plasmids#P2 P2]: BBa_J61002 (plasmid backbone with RFP) by XS
 +
*[https://2009.igem.org/Team:Paris/Freezer_Plasmids#P7 P7]: Bba_R0040 (ptet) by XP
 +
Digestion during 2h at 37°C.
 +
</div>
 +
<div class="guillaume">
 +
Gel migration
 +
</div>
 +
<div class="experience">
 +
*Gel migration for [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P1 P1]: pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P2 P2]: BBa_J61002 (plasmid backbone with RFP), [https://2009.igem.org/Team:Paris/Freezer_Plasmids#P7 P7]: BBa_R0040 (ptet)
 +
*[1kb|P1|P2|P7|100bp|/]
 +
 
 +
<center>[[Image:Wikidigestion060809.png]]</center>
 +
 
 +
*Result:
 +
**pSB2K3: wrong bande. Try again...
 +
**plasmid backbone with RFP: Wrong enzymes?!
 +
**ptet: good but the 1500bp band must not be there!
 +
</div>
 +
<div class="guillaume">
 +
ON culture
 +
</div>
 +
<div class="experience">
 +
*ON culture for [https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S8 S8]: pSB2K3, [https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S29 S29]: pSB1A3, [https://2009.igem.org/Team:Paris/Freezer_Strains_(Glycerol_stock)#S30 S30]: BBa_K136050 (RBS tetR) in order to done new miniprep
 +
</div>
 +
<div class="guillaume">
 +
Transformation
 +
</div>
 +
<div class="experience">
 +
*Resuspention of biobrick BBa_J2479 (1L7)
 +
*Transformation in competent DH5&alpha;
 +
</div>
<html>
<html>
</div>
</div>
Line 77: Line 110:
</html>
</html>
-
==To do list==
+
===To do list===
{|
{|
|- style="background: #ccccff; text-align: center;"
|- style="background: #ccccff; text-align: center;"
Line 115: Line 148:
|Sylvain
|Sylvain
|if(!oligo){return Maltose;} return PCR;
|if(!oligo){return Maltose;} return PCR;
-
|- style="background: #009900;text-align: center;"
+
|- style="background:green; text-align: center;"
|Guillaume
|Guillaume
-
|??
+
|Lab
|}
|}
 +
 +
 +
{{Paris2009_Calendar_Link|5_August_2009|7_August_2009}}

Latest revision as of 09:22, 13 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 6th

Lab work

Microscope

FM4-64 dying MG4 and Kayo delete Tol R

protocol n°2 (05/08/09) but using PBS

->nearly the same as MgSO4

protocol n°2 (05/08/09)but 1ml of cells and 1µl of dye, one with MgSO4 and one with PBS

->not enough dye

Molecular Biology

Gel Migration

  • [Ladder 1kb|A1|nothing|A2|nothing|A3]using 1% agarose during 30min
    • A1 = Tg3p = 274bp (utiliser le ladder 100bp)
    • A2 = TE3 = 1036bp
    • A3 = ClyA (with RBS and poly G linker = 987bp


KR000634.JPGA3 cut.JPG


  • A1 and A2 didn't work we don't see it on the gel because it's not in E.Coli.
  • A3 didn't migrate very well
    • [Ladder 1kb|nothing|A5|nothing|A5|nothing] using 1% agarose during 30min
    • A5 = TolRII = 279bp


KR000637.JPGA5 coupe.JPG

Purification and Digestion

  • Purification of A3, A4 directly in micro-column, A5, A6 using gel then micro-column
  • Digestion with XbaI and PSTI

New Miniprep

  • New Miniprep for P1: pSB2K3, P13: pSB1A3, P14: BBa_K136050 (RBS tetR)
  • Result: Just good for trash. Make new ON culture.

Digestion by XbaI/PstI (XP) or XbaI/SpeI (XS)

  • P1: pSB2K3 by XP
  • P2: BBa_J61002 (plasmid backbone with RFP) by XS
  • P7: Bba_R0040 (ptet) by XP

Digestion during 2h at 37°C.

Gel migration

  • Gel migration for P1: pSB2K3, P2: BBa_J61002 (plasmid backbone with RFP), P7: BBa_R0040 (ptet)
  • [1kb|P1|P2|P7|100bp|/]
Wikidigestion060809.png
  • Result:
    • pSB2K3: wrong bande. Try again...
    • plasmid backbone with RFP: Wrong enzymes?!
    • ptet: good but the 1500bp band must not be there!

ON culture

  • ON culture for S8: pSB2K3, S29: pSB1A3, S30: BBa_K136050 (RBS tetR) in order to done new miniprep

Transformation

  • Resuspention of biobrick BBa_J2479 (1L7)
  • Transformation in competent DH5α

To do list

Matricule TODO
Luc Labs
Romain Look for his feet
Charlotte Jun/Fos system (oligo + system)
Stoff DataBase / kitchen system ?
Chris modeling: model genetique network / next week lab planning / gillepsy
Lisa WTF !!!
Caroline lab / microscope
Souf lab / oligo / wiki
Vicard Lab : Gel :D
Pierre Tol/pal modeling
Sylvain if(!oligo){return Maltose;} return PCR;
Guillaume Lab


← Yesterday - Tomorrow →

Retrieved from "http://2009.igem.org/Team:Paris/6_August_2009"