Difference between revisions of "Team:Paris Saclay/Notebook/July/9"
(→Plasmid Rehydratation :) |
(→Wednesday 8th July) |
||
| Line 30: | Line 30: | ||
Plasmid with promotor: J23101 | Plasmid with promotor: J23101 | ||
| − | + | * 10µL of our plasmid with promotor | |
| − | + | * 1µL SpeI | |
| − | + | * 1µL PstI | |
| − | + | * 2µL buffer 10x FD | |
Plasmid with gene: COO40 and K115017 | Plasmid with gene: COO40 and K115017 | ||
| − | + | * 10µL of our plasmid with gene | |
| − | + | * 1µL XbaI | |
| − | + | * 1µL PstI | |
| − | + | * 2µL buffer 10x FD | |
After this step, we separate the sequence we need from the sequence we don't with electrophoresis | After this step, we separate the sequence we need from the sequence we don't with electrophoresis | ||
| Line 46: | Line 46: | ||
''by Pauline and Audrey'' | ''by Pauline and Audrey'' | ||
| − | Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0, | + | Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET |
Migration 0,06A 80V | Migration 0,06A 80V | ||
| Line 52: | Line 52: | ||
''by Pauline and Audrey'' | ''by Pauline and Audrey'' | ||
We quantify with the electrophoresis gel: | We quantify with the electrophoresis gel: | ||
| − | C0040: 10ng/ | + | * C0040: 10ng/µL |
| − | J23101: 20ng/ | + | * J23101: 20ng/µL |
| − | K115017: we don't see enough of fluorescent to quantify the biobrick. We decide to amplify this one by PCR. | + | * K115017: we don't see enough of fluorescent to quantify the biobrick. We decide to amplify this one by PCR. |
'''Members present:''' | '''Members present:''' | ||
* Instructors and advisors: Alice. | * Instructors and advisors: Alice. | ||
* Students: Johan, Seong Koo, Audrey, Coralie, Pauline | * Students: Johan, Seong Koo, Audrey, Coralie, Pauline | ||
Revision as of 10:45, 15 July 2015
Contents
Wednesday 8th July
Lab Work
Transformation
by Coralie
Ligation product:
- J23101 + I13504
- J23106 + I13504
- J23117 + I13504
On LB + Ampicillin 100ug/mL Incubation ON, 37°C
"by Johan and Seong Koo"
R0051 On LB + Ampicillin 100ug/mL Incubation ON, 37°C
Plasmid Rehydratation :
by Johan and Audrey
Biobricks:
- S03518
- B0030
- B0015
- K1399005
Digestion
by Pauline and Audrey
Plasmid with promotor: J23101
- 10µL of our plasmid with promotor
- 1µL SpeI
- 1µL PstI
- 2µL buffer 10x FD
Plasmid with gene: COO40 and K115017
- 10µL of our plasmid with gene
- 1µL XbaI
- 1µL PstI
- 2µL buffer 10x FD
After this step, we separate the sequence we need from the sequence we don't with electrophoresis
Electrophoresis
by Pauline and Audrey
Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET Migration 0,06A 80V
Quantification
by Pauline and Audrey We quantify with the electrophoresis gel:
- C0040: 10ng/µL
- J23101: 20ng/µL
- K115017: we don't see enough of fluorescent to quantify the biobrick. We decide to amplify this one by PCR.
Members present:
- Instructors and advisors: Alice.
- Students: Johan, Seong Koo, Audrey, Coralie, Pauline