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Team:Paris Saclay/Notebook/August/1
From 2014.igem.org
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=Friday 1st August= | =Friday 1st August= | ||
==Lab work== | ==Lab work== | ||
| - | === | + | ===The frame coli Odor free=== |
====Stria on LB dishes==== | ====Stria on LB dishes==== | ||
''by Romain'' | ''by Romain'' | ||
| Line 8: | Line 9: | ||
Strains used: MG1655 and MG1655Z1, transformed the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30 30th July], striated on 4 LB dishes per strain. | Strains used: MG1655 and MG1655Z1, transformed the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30 30th July], striated on 4 LB dishes per strain. | ||
| - | === | + | ===Salicylate Inducible Suppressing System=== |
====Liquid Culture==== | ====Liquid Culture==== | ||
''by Fabio'' | ''by Fabio'' | ||
| Line 14: | Line 15: | ||
8 liquid cultures with 3ml of LB and 3µl of amp (at 10am - 8°C), all from the colonies made the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#Bacterial_Culture 31st July]. The colonies will be put at 37°C on sunday 3rd in order to have grown colonies on monday. | 8 liquid cultures with 3ml of LB and 3µl of amp (at 10am - 8°C), all from the colonies made the [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#Bacterial_Culture 31st July]. The colonies will be put at 37°C on sunday 3rd in order to have grown colonies on monday. | ||
| - | === | + | ===Lemon scent=== |
====Results of the Transformation of electrocompetent cells==== | ====Results of the Transformation of electrocompetent cells==== | ||
''by Arnaud & Terry'' | ''by Arnaud & Terry'' | ||
| Line 26: | Line 27: | ||
*Failure, nothing has grown on the dishes which contain DY330 with pJBE1 | *Failure, nothing has grown on the dishes which contain DY330 with pJBE1 | ||
| + | ====New Transformation of electrocompetent cells==== | ||
| + | ''by Terry'' | ||
| + | |||
| + | Strain used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli) | ||
| + | |||
| + | Make 3 électroporations in cold electroporation cuvettes: | ||
| + | |||
| + | *A control cuvette(without DNA): 50µl of DY330. | ||
| + | *A second cuvette: 50µl of DY330 culture + 2µl of pJBEI-6409 plasmid. | ||
| + | *A third cuvette: 50µl of DY330 culture + 0.5µl of pJBEI-6409 plasmid. | ||
| + | |||
| + | Electroporation : 2500V, 132W, 40µF. | ||
| + | |||
| + | After that, add 1ml of cold LB in each cuvette and transfer in tubes to incubate during 1 to 2h at 30°C. | ||
| + | |||
| + | Spread on 7 dishes LB + Cm: | ||
| + | |||
| + | *100µl of control (without plasmid) | ||
| + | *50µl of transformed DY330 with 0.5µl of pJBEI-6409 | ||
| + | *100µl of transformed DY330 with 0.5µl of pJBEI-6409 | ||
| + | *The rest of transformed DY330 with 0.5µl pJBEI-6409 | ||
| + | *50µl of transformed DY330 with 2µl of pJBEI-6409 | ||
| + | *100µl of transformed DY330 with 2µl of pJBEI-6409 | ||
| + | *The rest of transformed DY330 with 2µl pJBEI-6409 | ||
| + | |||
| + | Incubate for 2 days at 30°C. | ||
| + | |||
| + | |||
| + | ==Photo of the Day== | ||
| + | [[File:Paris Saclay 1_august.jpg|600px|center]] | ||
'''Members there''': | '''Members there''': | ||
| Line 31: | Line 62: | ||
* Students: Arnaud, Fabio, Romain, Sean and Terry. | * Students: Arnaud, Fabio, Romain, Sean and Terry. | ||
| - | + | {{Team:Paris_Saclay/notebook_footer}} | |
Latest revision as of 14:23, 14 October 2014
Contents |
Friday 1st August
Lab work
The frame coli Odor free
Stria on LB dishes
by Romain
Strains used: MG1655 and MG1655Z1, transformed the 30th July, striated on 4 LB dishes per strain.
Salicylate Inducible Suppressing System
Liquid Culture
by Fabio
8 liquid cultures with 3ml of LB and 3µl of amp (at 10am - 8°C), all from the colonies made the 31st July. The colonies will be put at 37°C on sunday 3rd in order to have grown colonies on monday.
Lemon scent
Results of the Transformation of electrocompetent cells
by Arnaud & Terry
Transformation of electrocompetent cells made the July 29th. Strains used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)
Results:
- Nothing has grown on the control dishes.
- Failure, nothing has grown on the dishes which contain DY330 with pJBE1
New Transformation of electrocompetent cells
by Terry
Strain used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)
Make 3 électroporations in cold electroporation cuvettes:
- A control cuvette(without DNA): 50µl of DY330.
- A second cuvette: 50µl of DY330 culture + 2µl of pJBEI-6409 plasmid.
- A third cuvette: 50µl of DY330 culture + 0.5µl of pJBEI-6409 plasmid.
Electroporation : 2500V, 132W, 40µF.
After that, add 1ml of cold LB in each cuvette and transfer in tubes to incubate during 1 to 2h at 30°C.
Spread on 7 dishes LB + Cm:
- 100µl of control (without plasmid)
- 50µl of transformed DY330 with 0.5µl of pJBEI-6409
- 100µl of transformed DY330 with 0.5µl of pJBEI-6409
- The rest of transformed DY330 with 0.5µl pJBEI-6409
- 50µl of transformed DY330 with 2µl of pJBEI-6409
- 100µl of transformed DY330 with 2µl of pJBEI-6409
- The rest of transformed DY330 with 2µl pJBEI-6409
Incubate for 2 days at 30°C.
Photo of the Day
Members there:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Arnaud, Fabio, Romain, Sean and Terry.
