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Team:Paris Saclay/Notebook/August/6
From 2014.igem.org
Contents |
Wednesday 6th August
Lab work
A - The chassis coli Odor free
Final stock of MG1655
by Romain
2 final stocks of the MG1655 made with 1ml of strain and 250µl glycerol 87%, in -80°C.
C - Salicylate Inducible Suppressing System
Plasmid DNA Purification
by Eugene and Fabio
- BBa_K1372000 Cl.1
- BBa_K1372000 Cl.2
From Liquid Culture made the 5th August
Digestion
by Eugene and Fabio
In order to place the BBa_B0015 just after the BBa_K1372000, we will start the Assembly procedure again.
- BioBrick BBa_K1372000 (Salicylate promoter + NahR + RNA suppressor) as Part A
- BioBrick BBa_B0015 (Double terminator) as Part B
The enzymes used were:
- EcoRI and SpeI for BBa_K1372000
- EcoRI and XbaI for BBa_B0015
TODO: Illustration of the process
BioBrick Assembly - Digest Reaction Protocol
Segregate Process by Electrophoresis
by Eugene and Fabio
- BBa_K1372000 digested by EcoRI and SpeI
- BBa_B0015 digested by EcoRI and XbaI
Results:
In progress
BioBrick Assembly - Segregate Process Protocol
D - Lemon scent
PCR Cleanup
by Pierre & Sean
BioBricks used: BBa_K762100 and BBa_K517003 (PCR perfomed on the 4th August)
Gel electrophoresis
by Melanie
Plasmid used: p cola
Results inconclusive. Wrong buffer was used to prepare PCR.
PCR digestion
by Pierre & Sean
Protocol
For each BioBrick (Bba_K517003 and BBa_K762100)mix the following into a microcentrifuge tube.
| component | volume |
|---|---|
| H2O | 5.5μl |
| PacI | .5μl |
| Fast Digest buffer | 4μl |
| Cleaned-up PCR content | 30μl |
Then bain-marie at 37°C for one hour. Finally conduct PCR cleanup for both BioBricks, with 15µl of NE buffer.
PCR pCola plasmide (Geranyl Synthase)
by melanie
amplification of geranyl Synthase
Gel electrophoresis
(with Melanie)
Samples used: BBa_K517003 (2µl), BBa_K762100 (2µl), p cola(12µl)
Bibliography
By Hoang Vu
Still working on my mission: find more informations about limonene, pinene and citral A and B.I got now a couple of very interessant reviews about limonene and pinene.
Members present:
- Instructors and advisors: Alice.
- Students: Eugene, Fabio, Hoang Vu, Juliette, Melanie, Pierre, Romain, Sean and Terry.
