ULB/13 August 2009

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(New page: == August 13, 2009 == <div style='color:grey;'> ==Manipulation on plasmides== *Les bactéries qui ont reçu le plasmide avec RBS (BBa_B0034) ont bien poussé. *La ligation du promoteur-RBS...)
 
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== August 13, 2009 ==
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==Manipulation on plasmides==
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*Les bactéries qui ont reçu le plasmide avec RBS (BBa_B0034) ont bien poussé.
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*La ligation du promoteur-RBS-Plasmide (résistant au chloramphénicol) n’a pas fonctionné : la raison pourrait être protocole l’utilisation  du manuel d’assemblage, qui nous disait de laisser liguer 10 minutes à température ambiante, puis de désactiver les enzymes alors que la notice de la T4 ligase, conseil de la laisser 24h.
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*Vérification que les bricks sont présent dans le milieu dans nos extractions : Digestion du promoteur, promoteur+RBS, RBS+RFP et le terminateur par les enzymes de restrictions EcoRI et Spel pour les 2 premiers et Xbal et Pstl pour les autres.
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*Nous avons fait une PCR avec les amorces reçues pour ccdA programme à 55°C puis migration sur gel
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==Results==
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[[Image:13_8.jpg| les puits du dessus : Promoteur1, promoteur2, RBS+RFP1, RBS+RFP2, promoteur+RBS1, promoteur+RBS2, terminateur1, terminateur2.les puits du dessus : les résultats des migations des PCR]]
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/* Design for https://2009.igem.org/Team:ULB-Brussels
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    Université Libre de Bruxelles
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*[[Vérification des bricks]]:4 Adn amorces « normales », blanc amorces « normales » (problème ???), 4 ADN amorces inverses, blanc amorces inverses.
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html body {
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*[[PCR]]: tous les tubes ont réussi sauf que le  tube du blanc n’est pas à sa place (4eme position à la place de 6eme).
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|-
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|-
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!align="right"|{{#calendar: title=ULB |year=2009 | month=09}}
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|}
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==Manipulation on plasmides==
 +
*Bacteria with the plasmid with RBS (BBa_B0034) growed.
 +
*The ligation of the promoter-RBS-plasmid (chloramphenicol resistant) didn't work: the reason could be the use of the protocol of manual assembly, who told us to leave league 10 minutes at room temperature, then deactivate the enzymes, insted of 24 hours by the T4 ligase guide.
 +
*Verify that the bricks are now in the medium of our extractions: Digestion of the promoter and promoter + RBS by restriction enzymes EcoRI and Spel, RBS + RFP and the terminator by XbaI and PstI.
 +
*We made a PCR with primers for ccdA program received at 55 ° C then a gel migration.
 +
 +
==Results==
 +
 +
[[Image:13_8.jpg| les puits du dessus  : Promoteur1, promoteur2, RBS+RFP1, RBS+RFP2, promoteur+RBS1, promoteur+RBS2, terminateur1, terminateur2.les puits du dessus : les résultats des migations des PCR]]
 +
*'''Verification of bricks''':4 DNA primers "normal", white primers "normal", 4 DNA primers reverse, white reverse primers.
 +
* '''PCR''': all tubes have succeeded except the one control sample which is out of place (4th position instead of 6th).
</div>
</div>

Latest revision as of 21:13, 21 October 2009

iGEM Team:ULB-Brussels Wiki


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


Manipulation on plasmides

  • Bacteria with the plasmid with RBS (BBa_B0034) growed.
  • The ligation of the promoter-RBS-plasmid (chloramphenicol resistant) didn't work: the reason could be the use of the protocol of manual assembly, who told us to leave league 10 minutes at room temperature, then deactivate the enzymes, insted of 24 hours by the T4 ligase guide.
  • Verify that the bricks are now in the medium of our extractions: Digestion of the promoter and promoter + RBS by restriction enzymes EcoRI and Spel, RBS + RFP and the terminator by XbaI and PstI.
  • We made a PCR with primers for ccdA program received at 55 ° C then a gel migration.

Results

les puits du dessus  : Promoteur1, promoteur2, RBS+RFP1, RBS+RFP2, promoteur+RBS1, promoteur+RBS2, terminateur1, terminateur2.les puits du dessus : les résultats des migations des PCR

  • Verification of bricks:4 DNA primers "normal", white primers "normal", 4 DNA primers reverse, white reverse primers.
  • PCR: all tubes have succeeded except the one control sample which is out of place (4th position instead of 6th).

</div>

Retrieved from "http://2009.igem.org/ULB/13_August_2009"