Team:Paris Saclay/Notebook/July/22
From 2014.igem.org
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*Make extraction of 5ml of plasmid [https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue Protocol] | *Make extraction of 5ml of plasmid [https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue Protocol] | ||
- | Step 3 - When the culture | + | Step 3 - When the culture OD<sub>650</sub> = 0,6 (Step 1) |
'''Members there''': | '''Members there''': |
Revision as of 20:21, 23 July 2014
Contents |
Tuesday 22st July
Lab Work
1 - Samples for stock
by Fabio
We collected 1ml of each sample, added 240µl of glycerol (87%) and stored at -80°C.
- BBa_K1033902
- BBa_K1033905
- BBa_K1033910
- BBa_K1033913
- BBa_K1033922
- BBa_K1033925
- BBa_K1033927
- BBa_K1033929
- BBa_K103921
- BBa_J45017
- BBa_K731201
- BBa_K762100
from Liquide Bacterial Cultures transformed the 21st July
2 - Electrophoresis
by Arnaud (Process B), Fabio (gel), Marie and Romain (Process B)
We used 2µL of DNA of the following Biobricks in a 1% Agarose Gel.
Process A
Process B
Results:
- A:
- B:
- C:
3 - Bacterial Culture
by Fabio and Marie
One liquid culture with 5ml of LB (37°C - 150 rpm), IL1403 from the concentrated colony.
4 - Plasmid DNA extraction
by Sean and Terry
Plasmids used: cf part 1
Note: we accidentally used a buffer without 10% ethanol. After removal of the buffer, we repeated the step with a correct version. This may have affected the electrophoresis which followed.
C - Lemon scent
PCR Targeting
by Romain
Strains used: DY330. Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)
Step 1 - Dilution of 300µl of bacterial culture in 30ml of LB + 30µl Cm at 30°C.
Step 2 - With the pre-culture:
- make glycerol stock: 1ml bacterial culture with 240µl glycerol 87%.
- Make extraction of 5ml of plasmid Protocol
Step 3 - When the culture OD650 = 0,6 (Step 1)
Members there:
- Instructors and advisors: Solenne.
- Students: Arnaud, Fabio, Juliette, Mathieu, Marie, Pierre, Romain, Sean and Terry.