Team:Paris Saclay/Notebook/July/5

From 2013.igem.org

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{{Team:Paris_Saclay/incl_debut_generique}}
 
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='''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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=='''Lab work'''==
 
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*'''A.aero/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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<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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<br>
 
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{{Team:Paris_Saclay/incl_fin}}
 
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By using NanoDrop, we found some errors which could be due to device malfunctional issue.
By using NanoDrop, we found some errors which could be due to device malfunctional issue.
|}
|}
 +
 +
 +
 +
<u>Electrophoresis band size estimation</u><br>
 +
<p>We used Clonemanager for band size estimation:</p>
 +
<br>
 +
{| border="1" align="center"
 +
|-
 +
|Molecule
 +
|enzymes(Buffer)
 +
|Size
 +
|-
 +
|AmilCP clone
 +
|Nod I(Orange)
 +
|2046+693bp
 +
|-
 +
|AmilCP clone
 +
|XHOI(Red)
 +
|1842+892bp
 +
|-
 +
|BBa_K1155000
 +
|XHOI(Red)
 +
|1289+892bp
 +
|-
 +
|BBa_K1155000
 +
|ECoR I(Orange)
 +
|2151bp
 +
|}<br>
 +
 +
 +
 +
 +
 +
 +
 +
 +
 +
<br>
 +
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{| border="1" align="center"
{| border="1" align="center"

Revision as of 17:42, 22 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : August 23

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 : ...

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

2 - Extraction of Bba_K1155000 and Bba_K592009 from DH5α

Abdou

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

Protocol : Hight copy plamid extraction

Nanodrop :

  • Bba_K1155000P : 761ng/µl
  • Bba_K592009 : 229ng/µl

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


Electrophoresis band size estimation

We used Clonemanager for band size estimation:


Molecule enzymes(Buffer) Size
AmilCP clone Nod I(Orange) 2046+693bp
AmilCP clone XHOI(Red) 1842+892bp
BBa_K1155000 XHOI(Red) 1289+892bp
BBa_K1155000 ECoR I(Orange) 2151bp








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