Difference between revisions of "Team:Paris Saclay/Notebook/July/9"
m |
(→Electrophoresis) |
||
(5 intermediate revisions by 2 users not shown) | |||
Line 9: | Line 9: | ||
* BBa_J23106 + BBa_I13504 | * BBa_J23106 + BBa_I13504 | ||
* BBa_J23117 + BBa_I13504 | * BBa_J23117 + BBa_I13504 | ||
− | On LB + | + | On LB + Chloramphenicol 20ug/mL. |
Incubation ON, 37°C | Incubation ON, 37°C | ||
Line 46: | Line 46: | ||
Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET | Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET | ||
− | Migration 0,06A 80V | + | Migration 0,06A 80V |
+ | |||
+ | [[File:ParisSaclay_09072015_-_Quantification.jpg|300px|center]] | ||
+ | <html><i><p>Quantification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_C0040, 3. BBa_K115017, 4. BBa_J23101, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty, 11. Empty, 12. Empty</i></p></i></html> | ||
===Quantification=== | ===Quantification=== |
Latest revision as of 21:14, 18 September 2015
Contents
Thursday 9th July
Lab Work
Transformation
by Coralie
Ligation product:
- BBa_J23101 + BBa_I13504
- BBa_J23106 + BBa_I13504
- BBa_J23117 + BBa_I13504
On LB + Chloramphenicol 20ug/mL. Incubation ON, 37°C
by Johan and Seong Ko
- BBa_R0051
On LB + Ampicillin 100ug/mL. Incubation ON, 37°C
Plasmid Rehydratation
by Johan and Audrey
- BBa_S03518
- BBa_B0030
- BBa_B0015
- BBa_K1399005
Digestion
by Pauline and Audrey
Plasmid with promotor: BBa_J23101
- 10µL of our plasmid with promotor
- 1µL SpeI
- 1µL PstI
- 2µL buffer 10x FD
Plasmid with gene: BBa_COO40 and BBa_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, from left to right: 1. DNA Ladder, 2. BBa_C0040, 3. BBa_K115017, 4. BBa_J23101, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty, 11. Empty, 12. Empty
Quantification
by Pauline and Audrey
Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET Migration 0,06A 80V We quantify with the electrophoresis gel:
- BBa_C0040: 10ng/µL
- BBa_J23101: 20ng/µL
- BBa_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