Team:Paris Saclay/Notebook/July/5

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=='''Summary:'''==
=='''Summary:'''==
For fnr promoter regulator system:
For fnr promoter regulator system:
-
*Stocked 2 samples for each colonies which contained AmilCP or BBa_K1155000.
+
*Stocked 2 samples for each 2 colonies which contained AmilCP or BBa_K1155000.
-
*Performed PCR for amilCP and fnr from the medium seeded yesterday.
+
*Performed restriction digest for amilCP and fnr from the medium seeded yesterday.
*Verification DNA quantity by NanoDrop.
*Verification DNA quantity by NanoDrop.
*Performed electrophoresis and photo interpretation .
*Performed electrophoresis and photo interpretation .
For PCBs sensor system:
For PCBs sensor system:
-
*Stocked 2 samples of pseudomonas KE707.
+
*Stocked 2 samples of pseudomonas KE707. and performed a PCR
*Received Primer BphA1P_up/down and BphR2_up/down.
*Received Primer BphA1P_up/down and BphR2_up/down.
=='''Lab work'''==
=='''Lab work'''==
-
constructing
+
 
 +
*'''A.aero/anaerobic regulation system'''
 +
**''2.BioBrick RBS+LacZ+terminator in plasmid PSB1C3 ''
 +
**''BioBrick RBS+amilCP+terminator in plasmid PSB1C3 ''
 +
 
 +
<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>
 +
 
 +
<u>DNA quantification</u>
 +
<br>
 +
Result(average):
 +
*fnr promoter : 761ng/µl
 +
*AmilCP : 229ng/µl
 +
<p>By using NanoDrop, we found some erreurs which could be due to device malfunctional issue.</p><br>
 +
 
 +
<u>Electrophoresis</u><br>
 +
{| align="center"
 +
| style="width:350px;border:1px solid black;" | [[]]
 +
| style="width:350px;border:1px solid black;" |
 +
*Well 1,2 : Not I
 +
*Well 3,4 : Hpa I
 +
*Well 5.6 : Mlu I
 +
*gel 0.8%
 +
|}<br>
 +
 
 +
 
 +
*'''PCBs sensor system'''
 +
**''1.BioBrick BphR2(regulator) in plasmid PSB1C3''
 +
 
 +
 
 +
 
 +
 
 +
 
<br>
<br>
{| border="1" align="center"
{| border="1" align="center"

Revision as of 00:23, 19 September 2013

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Notebook : July 5

Summary:

For fnr promoter regulator system:

  • Stocked 2 samples for each 2 colonies which contained AmilCP or BBa_K1155000.
  • Performed restriction digest for amilCP and fnr from the medium seeded yesterday.
  • Verification DNA quantity by NanoDrop.
  • Performed electrophoresis and photo interpretation .

For PCBs sensor system:

  • Stocked 2 samples of pseudomonas KE707. and performed a PCR
  • Received Primer BphA1P_up/down and BphR2_up/down.

Lab work

  • A.aero/anaerobic regulation system
    • 2.BioBrick RBS+LacZ+terminator in plasmid PSB1C3
    • BioBrick RBS+amilCP+terminator in plasmid PSB1C3

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

DNA quantification
Result(average):

  • fnr promoter : 761ng/µl
  • AmilCP : 229ng/µl

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


Electrophoresis

[[]]
  • Well 1,2 : Not I
  • Well 3,4 : Hpa I
  • Well 5.6 : Mlu I
  • gel 0.8%


  • PCBs sensor system
    • 1.BioBrick BphR2(regulator) in plasmid PSB1C3




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