Team:Paris Saclay/Notebook/July/5

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{{Team:Paris_Saclay/incl_debut_generique}}
{{Team:Paris_Saclay/incl_debut_generique}}
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='''Notebook : July 5'''=
='''Notebook : July 5'''=
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=='''Summary:'''==
 
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For fnr promoter regulator system:
 
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*Stocked 2 samples for each 2 colonies which contained AmilCP or BBa_K1155000.
 
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*Performed restriction digest for amilCP and fnr from the medium seeded yesterday.
 
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*Verification DNA quantity by NanoDrop.
 
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*Performed electrophoresis and photo interpretation .
 
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For PCBs sensor system:
 
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*Stocked 2 samples of pseudomonas KE707. and performed a PCR
 
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*Received Primer BphA1P_up/down and BphR2_up/down.
 
=='''Lab work'''==
=='''Lab work'''==
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*'''A.aero/anaerobic regulation system'''
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==='''A - Aerobic/Anaerobic regulation system'''===
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**''2.BioBrick RBS+LacZ+terminator in plasmid PSB1C3 ''
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**''BioBrick RBS+amilCP+terminator in plasmid PSB1C3 ''
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<u>Electrophoresis band size estimation</u><br>
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===='''Objective : obtaining BBa_K1155000'''====
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<p>We used Clonemanager for band size estimation:</p>
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<br>
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{| border="1" align="center"
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|-
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|Molecule
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|enzymes(Buffer)
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|Size
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|-
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|AmilCP clone
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|Nod I(Orange)
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|2046+693bp
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|-
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|AmilCP clone
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|XHOI(Red)
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|1842+892bp
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|-
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|BBa_K1155000
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|XHOI(Red)
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|1289+892bp
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|-
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|BBa_K1155000
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|ECoR I(Orange)
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|2151bp
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|}<br>
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<u>DNA quantification</u>
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===='''1 - Electrophoresis of the digestion of BBa_K1155000 by NotI, HpaI, MluI'''====
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<br>
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Result(average):
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Sheng, Zhou
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*fnr promoter : 761ng/µl
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*AmilCP : 229ng/µl
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<p>By using NanoDrop, we found some erreurs which could be due to device malfunctional issue.</p><br>
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<u>Electrophoresis</u><br>
 
{| align="center"
{| align="center"
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| style="width:350px;border:1px solid black;" | [[[[File:PCRPS050813.jpg|right|350px]]]]
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| style="width:350px;border:1px solid black;" | [[File:PCRPS050813.jpg|right|350px]]
| style="width:350px;border:1px solid black;" |
| style="width:350px;border:1px solid black;" |
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*Well 1,2 : Not I
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* Well 1, 2 : 5µL of BBa_K1155000 digested by Not I+1µL of 6X loading dye
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*Well 3,4 : Hpa I
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* Well 3, 4 : 5µL of BBa_K1155000 digested by Hpa I+1µL of 6X loading dye
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*Well 6,7 : Mlu I
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* Well 5 : 6µL of DNA Ladder
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*gel 0.8%
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* Well 6, 7 : 5µL of BBa_K1155000 digested by Mlu I+1µL of 6X loading dye
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|}<br>
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* Gel 0.8%
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|}
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Expected sizes :
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* BBa_K1155000 : 111bp
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{|
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| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
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We didn't obtain fragments at the right size. We will do digestions again.
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|}
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===='''2 - Extraction of BBa_K1155000 from DH5α'''====
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Abdou
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{|
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| style="border:1px solid black;padding:5px;background-color:#DE;" |
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Transformation of 07/04 works. We will extract DNA plasmid.
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|}
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Protocol : [[Team:Paris_Saclay/extraction|High copy plamid extraction]]
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Nanodrop :
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* BBa_K1155000 : 761ng/µl
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{|
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| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
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By using NanoDrop, we found some errors which could be due to device malfunctional issue.
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|}
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===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''====
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It seemed that we had made some terrible mistakes. We continued and did another digestion the next week.
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===='''1 - Extraction of BBa_K592009 and BBa_I732017 from DH5α'''====
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*'''PCBs sensor system'''
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Abdou
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**''1.BioBrick BphR2(regulator) in plasmid PSB1C3''
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{|
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| style="border:1px solid black;padding:5px;background-color:#DE;" |
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Transformation of 07/04 works. We will extract DNA plasmid.
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|}
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Protocol : [[Team:Paris_Saclay/extraction|High copy plamid extraction]]
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Nanodrop :
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* BBa_K592009 : 229ng/µl
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{|
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| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" |
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By using NanoDrop, we found some errors which could be due to device malfunctional issue.
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|}
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<br>
 
{| border="1" align="center"
{| border="1" align="center"
|[[Team:Paris Saclay/Notebook/July/4|<big>Previous day</big>]]
|[[Team:Paris Saclay/Notebook/July/4|<big>Previous day</big>]]

Latest revision as of 16:35, 4 October 2013

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Contents

Notebook : July 5

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155000

1 - Electrophoresis of the digestion of BBa_K1155000 by NotI, HpaI, MluI

Sheng, Zhou

PCRPS050813.jpg
  • Well 1, 2 : 5µL of BBa_K1155000 digested by Not I+1µL of 6X loading dye
  • Well 3, 4 : 5µL of BBa_K1155000 digested by Hpa I+1µL of 6X loading dye
  • Well 5 : 6µL of DNA Ladder
  • Well 6, 7 : 5µL of BBa_K1155000 digested by Mlu I+1µL of 6X loading dye
  • Gel 0.8%

Expected sizes :

  • BBa_K1155000 : 111bp

We didn't obtain fragments at the right size. We will do digestions again.

2 - Extraction of BBa_K1155000 from DH5α

Abdou

Transformation of 07/04 works. We will extract DNA plasmid.

Protocol : High copy plamid extraction

Nanodrop :

  • BBa_K1155000 : 761ng/µl

By using NanoDrop, we found some errors which could be due to device malfunctional issue.

Objective : obtaining BBa_K1155003, BBa_K1155007

1 - Extraction of BBa_K592009 and BBa_I732017 from DH5α

Abdou

Transformation of 07/04 works. We will extract DNA plasmid.

Protocol : High copy plamid extraction

Nanodrop :

  • BBa_K592009 : 229ng/µl

By using NanoDrop, we found some errors which could be due to device malfunctional issue.


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