Team:Paris Saclay/Notebook/July/10

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{{Team:Paris_Saclay/incl_debut_generique}}
{{Team:Paris_Saclay/incl_debut_generique}}
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='''Notebook : July 10'''=
='''Notebook : July 10'''=
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==''Summary:''==
 
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For regulation system:
 
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*prepared the solution of BioBrick fnr repressor in PsB1C3 plasmid for DNA sequencing.
 
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*the terminator transformation of BBa_B0010 did not work yesterday, a second transformation had been done for it.
 
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*The transformation for RBS+LacZ+terminator plasmid into competent cells was performed.
 
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*after the transformation PSB3K3 plasmid in competent cells, these cells were cultured in a liquid nutritive medium.
 
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For PSBs sensor system:
 
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*the ligation products were transformed into competent cells and were cultured on solid medium with their specific antibiotics.
 
=='''Lab work'''==
=='''Lab work'''==
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A.aero/anaerobic regulation system:
 
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*BioBrick RBS+LacZ+terminator in plasmid PSB1C3
 
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*BioBrick RBS+amilCP+terminator in plasmid PSB1C3
 
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<u>Transformation</u><br>
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==='''A - Aerobic/Anaerobic regulation system'''===
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<p>Transformation for BBa_B0010 dit work, we observed 0 colonies on the Petri dish, We decided to redo another transformation</p><br>
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===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''====
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<p>Transformation for BBa_I732019(RBS+LacZ+terminator BBa_B0010):
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===='''1 - Transformation of BBa_I732019 in DH5α'''====
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:the construction of RBS+LacZ+terminator BBa_B0010 is already done and stocked in iGEM BioBrick bank named BBa_I732019 12G p4 kit 2012. So we just suspended the BioBrick with 10µl water, transformed them into 100µl competent cell.</p><br>
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<p>Cloning for plasmid PSB3K3:
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Sheng
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:results of transformation and cloning: 34 colonies grown.
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:We picked up 2 colonies, seeded them in liquid medium(LB+chloramphenicol), incubation at 37°C, 200rpm during one night.</p><br>
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Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
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===='''Objective : obtaining biobricks in pSB3K3'''====
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===='''1 - Mini and maxi preparation of the culture of BBa_J04450'''====
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{{Team:Paris_Saclay/incl_debut_generique}}
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='''Notebook : August 23'''=
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=='''Lab work'''==
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==='''A - Aerobic/Anaerobic regulation system'''===
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===='''Objective : obtaining Bba_K1155003, Bba_K1155007'''====
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===='''1 - Transformation of Bba_I732019 and Bba_B0010 in DH5α'''====
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Sheng
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{|
{|
| style="border:1px solid black;padding:5px;background-color:#DE;" |
| style="border:1px solid black;padding:5px;background-color:#DE;" |
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Transformation of Bba_B0010 of 07/09/13 did work. We will do it again.
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Transformation of BBa_J044500 of 07/09/13 didn't work. We obtain 34 colonies. We will extract DNA by doing mini and maxi preparations.
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Protocol : [[Team:Paris_Saclay/Protocols/Bacterial transformation|Bacterial transformation]]
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Protocol : [[Team:Paris_Saclay/mini_maxi|Mini and maxi preparation]]
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===='''Objective : obtaining biobricks in PSB3K3'''====
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===='''1 - '''====
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We let our preparations at 37°C, 200rpm with 5mL of LB and 5mL of kanamicine antibiotic.
==='''B - PCB sensor system'''===
==='''B - PCB sensor system'''===
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===='''Objective : obtaining Bba_K1155001, Bba_K1155002, BphR2 protein'''====
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===='''Objective : obtaining BBa_K1155001, BBa_K1155002, BphR2 protein'''====
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===='''1 - Transformation of ligation of PSB1C3 and BphA1 or BphR1 or BphR2'''====
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===='''1 - Transformation of ligation of pSB1C3 and BphA1 or BphR1 or BphR2'''====
Abdou
Abdou
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Protocol : [[Team:Paris_Saclay/Protocols/Bacterial transformation|Bacterial transformation]]
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Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]

Latest revision as of 16:36, 4 October 2013

Contents

Notebook : July 10

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155003, BBa_K1155007

1 - Transformation of BBa_I732019 in DH5α

Sheng

Protocol : Bacterial transformation

Objective : obtaining biobricks in pSB3K3

1 - Mini and maxi preparation of the culture of BBa_J04450

Abdou

Transformation of BBa_J044500 of 07/09/13 didn't work. We obtain 34 colonies. We will extract DNA by doing mini and maxi preparations.

Protocol : Mini and maxi preparation

We let our preparations at 37°C, 200rpm with 5mL of LB and 5mL of kanamicine antibiotic.


B - PCB sensor system

Objective : obtaining BBa_K1155001, BBa_K1155002, BphR2 protein

1 - Transformation of ligation of pSB1C3 and BphA1 or BphR1 or BphR2

Abdou

Protocol : Bacterial transformation


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