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Team:Paris Saclay/Notebook/July/16
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==='''A - Aerobic/Anaerobic regulation system'''=== | ==='''A - Aerobic/Anaerobic regulation system'''=== | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''==== |
| - | ===='''1 - Extraction of | + | ===='''1 - Extraction of BBa_B0015, BBa_B0017, BBa_I732019 from DH5α'''==== |
Zhou | Zhou | ||
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Protocol : [[Team:Paris_Saclay/extraction|High copy plamid extraction]] | Protocol : [[Team:Paris_Saclay/extraction|High copy plamid extraction]] | ||
| - | ===='''2 - Digestion of | + | ===='''2 - Digestion of BBa_B0015, BBa_B0017, BBa_I732019 by EcoRI/PstI'''==== |
Anaïs | Anaïs | ||
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===='''Objective : obtaining biobricks in PSB3K3'''==== | ===='''Objective : obtaining biobricks in PSB3K3'''==== | ||
| - | ===='''1 - Digestion of | + | ===='''1 - Digestion of pSB3K3 by EcoRI/PstI'''==== |
Anaïs | Anaïs | ||
| Line 40: | Line 40: | ||
* H2O : 11µL | * H2O : 11µL | ||
| - | We let our digestion 1h30 at 37°C. | + | We let our digestion 1h30 at 37°C.p |
| - | + | ===='''2 - Electrophoresis of the digestion of pSB3K3 by EcoRI/PstI'''==== | |
| - | ===='''2 - Electrophoresis of the digestion of | + | |
Sheng | Sheng | ||
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| style="width:350px;border:1px solid black;" |[[File:Psgel11607.jpg|500px]] | | style="width:350px;border:1px solid black;" |[[File:Psgel11607.jpg|500px]] | ||
| style="width:350px;border:1px solid black;vertical-align:top;" | | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
| - | * Well 1 : 5µL of | + | * Well 1 : 5µL of pSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye |
| - | * Well 2 : 5µL of | + | * Well 2 : 5µL of pSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye |
* Well 3 : 6µL of DNA ladder | * Well 3 : 6µL of DNA ladder | ||
| - | * Well 4 : 5µL of | + | * Well 4 : 5µL of pSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye |
| - | * Well 5 : 5µL of | + | * Well 5 : 5µL of pSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye |
* Gel : 1% | * Gel : 1% | ||
|} | |} | ||
Expected sizes : | Expected sizes : | ||
| - | * | + | * pSB3K3 : 2750bp |
{| | {| | ||
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|} | |} | ||
| - | ===='''3 - Gel purification of electrophoresis of the digestion of | + | ===='''3 - Gel purification of electrophoresis of the digestion of pSB3K3 by EcoRI/PstI'''==== |
Abdou | Abdou | ||
| Line 74: | Line 73: | ||
==='''B - PCB sensor system'''=== | ==='''B - PCB sensor system'''=== | ||
| - | ===='''Objective : obtaining | + | ===='''Objective : obtaining BBa_K1155001, BBa_K1155002, BphR2 in pSB1C3'''==== |
| - | ===='''1 - Electrophoresis of the digestion of | + | ===='''1 - Electrophoresis of the digestion of BBa_K1155001, BBa_K1155002, BphR2 in pSB1C3'''==== |
Anaïs, Zhou | Anaïs, Zhou | ||
| - | * | + | * BBa_K1155001 : |
{| | {| | ||
| - | | style="width:350px;border:1px solid black;" |[[File: | + | | style="width:350px;border:1px solid black;" |[[File:Psgel21607.jpg|300px]] |
| style="width:350px;border:1px solid black;vertical-align:top;" | | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
| - | * Well 1 to 5 : 2µL | + | * Well 1 to 5 : 2µL BBa_K1155001 digested by EcoRI/PstI+2µl of 6X loading dye |
| - | * Well 6 and 7 : 2µL | + | * Well 6 and 7 : 2µL BBa_K1155001 digested by SacII+2µl of 6X loading dye |
* Well 8 : 6µL DNA Ladder | * Well 8 : 6µL DNA Ladder | ||
* Gel : 1.2% | * Gel : 1.2% | ||
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Expected size : | Expected size : | ||
| - | * | + | * BBa_K1155001 digested by EcoRI/PstI : 2037bp + 333bp |
| - | * | + | * BBa_K1155001 digested by SacII : 2370bp |
{| | {| | ||
| style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | | style="border:1px solid black;padding:5px;background-color:#DEDEDE;" | | ||
| - | We obtain fragment at the right size. We will amplify | + | We obtain fragment at the right size. We will amplify BBa_K1155001. |
|} | |} | ||
| - | * | + | * BBa_K1155002 : |
{| | {| | ||
| - | | style="width:350px;border:1px solid black;" |[[File: | + | | style="width:350px;border:1px solid black;" |[[File:Psgel31607.jpg|500px]] |
| style="width:350px;border:1px solid black;vertical-align:top;" | | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
| - | * Well 1 to 8 : 2µL | + | * Well 1 to 8 : 2µL BBa_K1155002 digested by EcoRI/PstI+2µl of 6X loading dye |
* Well 9 : 6µL DNA Ladder | * Well 9 : 6µL DNA Ladder | ||
| - | * Well 10 to 17 : 2µL | + | * Well 10 to 17 : 2µL BBa_K1155002 digested by SacII+2µl of 6X loading dye |
* Gel : 1.5% | * Gel : 1.5% | ||
|} | |} | ||
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{| | {| | ||
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We didn't obtain fragments at the right size. We will do the ligation again. | We didn't obtain fragments at the right size. We will do the ligation again. | ||
|} | |} | ||
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{| | {| | ||
| - | | style="width:350px;border:1px solid black;" |[[File: | + | | style="width:350px;border:1px solid black;" |[[File:Psgel41607.jpg|500px]] |
| style="width:350px;border:1px solid black;vertical-align:top;" | | | style="width:350px;border:1px solid black;vertical-align:top;" | | ||
| - | * Well 1 to 8 : 2µL BphR2 in | + | * Well 1 to 8 : 2µL BphR2 in pSB1C3 digested by EcoRI/PstI+2µl of 6X loading dye |
* Well 9 : 6µL DNA Ladder | * Well 9 : 6µL DNA Ladder | ||
| - | * Well 10 to 17 : 2µL BphR2 in | + | * Well 10 to 17 : 2µL BphR2 in pSB1C3 digested by SacII+2µl of 6X loading dye |
* Gel : 1.5% | * Gel : 1.5% | ||
|} | |} | ||
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{| | {| | ||
Latest revision as of 00:02, 5 October 2013
Notebook : July 16
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining BBa_K1155003, BBa_K1155007
1 - Extraction of BBa_B0015, BBa_B0017, BBa_I732019 from DH5α
Zhou
Protocol : High copy plamid extraction
2 - Digestion of BBa_B0015, BBa_B0017, BBa_I732019 by EcoRI/PstI
Anaïs
- DNA : 5µL
- Buffer FD : 2µL
- EcoRI FD : 1µL
- PstI FD : 1µL
- H2O : 21µL
We let our digestion 1h30 at 37°C.
Objective : obtaining biobricks in PSB3K3
1 - Digestion of pSB3K3 by EcoRI/PstI
Anaïs
Used quantities :
- DNA : 5µL
- Buffer FD : 2µL
- EcoRI FD : 1µL
- PstI FD : 1µL
- H2O : 11µL
We let our digestion 1h30 at 37°C.p
2 - Electrophoresis of the digestion of pSB3K3 by EcoRI/PstI
Sheng
|
|
Expected sizes :
- pSB3K3 : 2750bp
|
We obtain fragments at the right size. |
3 - Gel purification of electrophoresis of the digestion of pSB3K3 by EcoRI/PstI
Abdou
Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
B - PCB sensor system
Objective : obtaining BBa_K1155001, BBa_K1155002, BphR2 in pSB1C3
1 - Electrophoresis of the digestion of BBa_K1155001, BBa_K1155002, BphR2 in pSB1C3
Anaïs, Zhou
- BBa_K1155001 :
|
|
Expected size :
- BBa_K1155001 digested by EcoRI/PstI : 2037bp + 333bp
- BBa_K1155001 digested by SacII : 2370bp
|
We obtain fragment at the right size. We will amplify BBa_K1155001. |
- BBa_K1155002 :
|
|
|
We didn't obtain fragments at the right size. We will do the ligation again. |
|
|
|
We didn't obtain fragments at the right size. After reading the sequence again, we find an digestion site in the middle of our biobrick. We will do a Gibson assembly to modify this site. |
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