From 2013.igem.org
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| | {{Team:Paris_Saclay/incl_debut_generique}} | | {{Team:Paris_Saclay/incl_debut_generique}} |
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| | ='''Notebook : July 8'''= | | ='''Notebook : July 8'''= |
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| - | ==''Summary:''==
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| - | FNR regulator system:
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| - | *performed another digestion for AmilCP, Pfnr plus sequence LacZ with Not I, Xho I, EcoR I and PST I.
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| - | *electrophoresis for those digest products
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| - | *enrichment culture for clone of the briobrick BBa_K1155000
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| - | *electrophoresis for extracted products of last friday
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| - | =='''Lab work'''==
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| - | Overnight incubation at 37°C with agitation.
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| - | <br>
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| - | The electrophoresis for the plasmids which we had extracted last week plus 3 other BioBrick promoter Bph R1, promoter Bph A1, Bph R2.
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| - | Volume added:
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| - | *Pfnr2: 10µl
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| - | *R1P: 10µl
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| - | *R2: 10µl
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| - | *A1P: 10µl
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| - | Migration at 100V for 25mins then continued till 45 minutes:
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| - | <br>
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| - | Results:
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| - | {| align="center"
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| - | | style="width:350px;border:1px solid black;" | [[File:PS08071345.jpg|right|350px]]
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| - | | style="width:350px;border:1px solid black;" |
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| - | *Well 1 : R2
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| - | *Well 2 : R1P
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| - | *Well 3 : A1P
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| - | *Well 4 : Pfnr
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| - | *gel 0.8%
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| - | |}<br>
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| - | The concentration of Pfnr2 was detected by NanoDrop :
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| - | *C = 1788.8 ng/µl
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| - | *260/280 = 2.08
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| - | We considered that our extraction of plasmid DNA was not successful, we need to perform another extraction DNA.
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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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| - | {| border="1" align="center"
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| - | |Molecule
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| - | |enzymes(Buffer)
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| - | |Size
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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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| - | {{Team:Paris_Saclay/incl_debut_generique}}
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| - | ='''Notebook : August 23'''=
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| | =='''Lab work'''== | | =='''Lab work'''== |
Revision as of 15:04, 26 September 2013
Notebook : July 8
Lab work
A - Aerobic/Anaerobic regulation system
Objective : Bba_K1155003, Bba_K1155007
1 - Digestion of Bba_K592009, Bba_K1155007 by NotI, XhoI, EcoRI, EcoRI/PstI
Abou, Anaïs, Sheng, Zhou
Used quantities :
- NotI :
- Bba_K592009 : 2µL
- Buffer orange : 2µL
- NotI : 0.5µL
- H2O : 15.5µL
- XhoI :
- Bba_K592009 : 2µL
- Buffer red : 2µL
- XhoI : 0.5µL
- H2O : 15.5µL
- EcoRI :
- Bba_K592009 : 2µL
- Buffer orange : 2µL
- EcoRI : 0.5µL
- H2O : 15.5µL
- EcoRI/PstI :
- Bba_I732017 : 2µL
- Buffer orange : 2µL
- EcoRI : 0.5µL
- PstI : 0.5µL
- H2O : 15µL
We let our digestion 1h30 at 37°C.
2 - Electrophoresis of the digestion of Bba_K592009, Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI
Abdou, Anaïs, Sheng, Zhou
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- Well 1 : 6µL DNA Ladder
- Well 2 : 5µL Bba_K592009 clone 1 digested by EcoRI+1µl of 6X loading dye
- Well 3 : 5µL Bba_K592009 clone 2 digested by EcoRI+1µl of 6X loading dye
- Well 4 : 5µL Bba_K592009 clone 1 digested by NotI+1µl of 6X loading dye
- Well 5 : 5µL Bba_K592009 clone 2 digested by NotI+1µl of 6X loading dye
- Well 6 : 5µL Bba_K592009 clone 1 digested by XhoI+1µl of 6X loading dye
- Well 7 : 5µL Bba_K592009 clone 2 digested by XhoI+1µl of 6X loading dye
- Well 8 : 5µL Bba_I732017 clone 1 digested by EcoRI/PstI+1µl of 6X loading dye
- Well 9 : 5µL Bba_I732017 clone 2 digested by EcoRI/PstI+1µl of 6X loading dye
- Well 10 : -
- Well 11 : -
- Well 12 : 6µL DNA Ladder
- Gel : 1%
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Expected sizes :
- Bba_K592009 digested by NotI : 2046bp + 693bp
- Bba_K592009 digested by XhoI : 1842bp + 892bp
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