Difference between revisions of "Team:Paris Saclay/Notebook/July/9"
(→Thursday 9th July) |
m |
||
Line 1: | Line 1: | ||
+ | {{Team:Paris_Saclay/notebook_header}} | ||
=Thursday 9th July= | =Thursday 9th July= | ||
==Lab Work== | ==Lab Work== | ||
− | + | ===Transformation=== | |
''by Coralie'' | ''by Coralie'' | ||
Ligation product: | Ligation product: | ||
− | * | + | * BBa_J23101 + BBa_I13504 |
− | * | + | * BBa_J23106 + BBa_I13504 |
− | * | + | * BBa_J23117 + BBa_I13504 |
− | On LB + Ampicillin 100ug/mL | + | On LB + Ampicillin 100ug/mL. |
Incubation ON, 37°C | Incubation ON, 37°C | ||
− | |||
''by Johan and Seong Ko'' | ''by Johan and Seong Ko'' | ||
− | + | * BBa_R0051 | |
− | + | On LB + Ampicillin 100ug/mL. | |
− | On LB + Ampicillin 100ug/mL | + | |
Incubation ON, 37°C | Incubation ON, 37°C | ||
− | + | ===Plasmid Rehydratation=== | |
''by Johan and Audrey'' | ''by Johan and Audrey'' | ||
− | + | * BBa_S03518 | |
− | * | + | * BBa_B0030 |
− | * | + | * BBa_B0015 |
− | * | + | * BBa_K1399005 |
− | * | + | |
− | + | ===Digestion=== | |
''by Pauline and Audrey'' | ''by Pauline and Audrey'' | ||
− | Plasmid with promotor: | + | Plasmid with promotor: BBa_J23101 |
* 10µL of our plasmid with promotor | * 10µL of our plasmid with promotor | ||
* 1µL SpeI | * 1µL SpeI | ||
Line 36: | Line 34: | ||
* 2µL buffer 10x FD | * 2µL buffer 10x FD | ||
− | Plasmid with gene: | + | Plasmid with gene: BBa_COO40 and BBa_K115017 |
* 10µL of our plasmid with gene | * 10µL of our plasmid with gene | ||
* 1µL XbaI | * 1µL XbaI | ||
Line 44: | Line 42: | ||
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 | ||
− | + | ===Electrophoresis=== | |
''by Pauline and Audrey'' | ''by Pauline and Audrey'' | ||
Line 50: | Line 48: | ||
Migration 0,06A 80V | Migration 0,06A 80V | ||
− | + | ===Quantification=== | |
''by Pauline and Audrey'' | ''by Pauline and Audrey'' | ||
Line 56: | Line 54: | ||
Migration 0,06A 80V | Migration 0,06A 80V | ||
We quantify with the electrophoresis gel: | 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:''' | '''Members present:''' | ||
* Instructors and advisors: Alice. | * Instructors and advisors: Alice. | ||
* Students: Johan, Seong Koo, Audrey, Coralie, Pauline | * Students: Johan, Seong Koo, Audrey, Coralie, Pauline | ||
+ | {{Team:Paris_Saclay/notebook_footer}} |
Revision as of 19:54, 21 July 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 + Ampicillin 100ug/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
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