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Team:Paris Saclay/Notebook/August/4
From 2014.igem.org
Contents |
Monday 4th August
Lab Work
A - The chassis coli Odor free
Striate on Dishes
by Juliette & Romain
We striated MG1655 and MG1655Z1 on LB and Kan dishes, from the striates prepared on LB dishes 1st August
C - Salicylate Inducible Suppressing System
Plasmid DNA Purification
by Eugene and Fabio
- Ligation of BBa_J61051 and BBa_K228001 made the 30th July
From Liquid Culture made the 1st August
Digestion to check
In progress
Electrophoresis
In progress
D - Lemon scent
Gel electrophoresis of BBa_K762100
by Sean
Samples used were prepared on the 30th July.
Results
From left to right: 5 µl of BBa_K762100 + "old" Phusion enzyme, 5 µl of BBa_K762100 + "new" Phusion enzyme
Electroporation
by Terry
Strain used: DY330 containing plasmid pJBEI6409.
We had to switch the Limonen Synthase cassette with the Apramycin resistance.
The culture has been placed at 42°C for 17 min then cooled 10 min before centrifugation.
Polymerase chain reaction
by Sean & Pierre
BBa_K517003
| component | volume |
|---|---|
| H2O | 35.5μl |
| Phusion buffer 5X | 10μl |
| dNTPs | 2μl |
| iPS68bis | 1μl |
| iPS69 | 1μl |
| DNA | 1μl |
| Phusion enzyme | 0.5μl |
BBa_K762100
For this BioBrick two tubes were prepared: one with undiluted DNA and one with DNA diluted 10-1.
| component | volume |
|---|---|
| H2O | 35.5μl |
| Phusion buffer 5X | 10μl |
| dNTPs | 2μl |
| iPS66 | 1μl |
| iPS67 | 1μl |
| DNA | 1μl |
| Phusion enzyme | 0.5μl |
Tubes were placed in PCR machine with the following parameters.
| Cycle step | Temperature | Time | Cycle |
|---|---|---|---|
|
Initial denaturation |
98°C |
1 min |
1 |
| Denaturation | 98°C | 15 s | 25 - 30 |
| Annealing | 52°C | 25 s | 25 - 30 |
| Extension | 72°C | 45 s | 25-30 |
| Final extension | 72°C | 10 min | 1 |
| Final extension | 8°C | hold | 1 |
Members there:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Eugene, Fabio, Hoang Vu, Juliette, Pierre, Romain, Sean and Terry.
