Team:Paris Saclay/Notebook/July/10

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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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<br>
 
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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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<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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<p>Transformation for BBa_I732019(RBS+LacZ+terminator BBa_B0010):
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==='''A - Aerobic/Anaerobic regulation system'''===
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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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===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''====
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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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B.PCBs sensor system:
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===='''1 - Transformation of BBa_I732019 in DH5α'''====
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:*Contruction for BioBrick promoter BphR1, BphR2, BphA1
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<u>Transformation and cloning</u>
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Sheng
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<p>From the ligation products which we performed yesterday, We inserted those BioBrick into competent cell( 5µl of ligation product with 50µl competent cell). Then cloning on Petri dish(LB with antibiotics chloamphénicol), incubation during 1 night at 37°C.</p>
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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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Abdou
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| style="border:1px solid black;padding:5px;background-color:#DE;" |
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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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|}
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Protocol : [[Team:Paris_Saclay/mini_maxi|Mini and maxi preparation]]
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We let our preparations at 37°C, 200rpm with 5mL of LB and 5mL of kanamicine antibiotic.
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==='''B - PCB sensor system'''===
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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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Abdou
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Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
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|[[Team:Paris Saclay/Notebook/July/9|<big>Previous day</big>]]
|[[Team:Paris Saclay/Notebook/July/9|<big>Previous day</big>]]

Latest revision as of 16:36, 4 October 2013

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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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