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Team:Paris Saclay/Notebook/August/13
From 2014.igem.org
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==Lab work== | ==Lab work== | ||
| - | === | + | ===Lemon scent=== |
| + | ====culture of pJBEI6409 bacteria==== | ||
| + | ''by Mélanie'' | ||
| + | |||
| + | culture in LB of some clones obtain yesterday | ||
====Gel electrophoresis of BBa_K762100==== | ====Gel electrophoresis of BBa_K762100==== | ||
''by Sean'' | ''by Sean'' | ||
| + | |||
| + | Samples used: 5µl from each tube prepared [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12#PCR_of_BBa_K762100 12th August] | ||
| + | |||
| + | Contents of the two tubes yielding the highest intensity were pooled. | ||
| + | |||
| + | |||
| + | ====PCR clean-up of BBa_K762100==== | ||
| + | ''by Sean'' | ||
| + | |||
| + | PCR clean-up was conducted on the contents of the two tubes pooled earlier. | ||
| + | |||
| + | |||
| + | ====Gel electrophoresis of BBa_K762100==== | ||
| + | ''by Sean'' | ||
| + | |||
| + | Sample used: the result of the PCR clean-up conducted today. | ||
| + | |||
| + | '''''Concentration of BBa_K762100 is very low — the band is only slightly visible.''''' | ||
| + | |||
====Cloning BBa_K517003 and p cola in pGEMTeasy==== | ====Cloning BBa_K517003 and p cola in pGEMTeasy==== | ||
| Line 80: | Line 103: | ||
We used 3 colony of bacteria transformed with PJBEI+Apra (11/08) to lauch 3 cultures | We used 3 colony of bacteria transformed with PJBEI+Apra (11/08) to lauch 3 cultures | ||
| - | + | -two cultures in 5mL LB+Apra (1/2000) | |
| - | + | -one culture in 20mL LB+Apra (1/2000) | |
37°C - 24h | 37°C - 24h | ||
| Line 95: | Line 118: | ||
The mold was not hermetically closed, the chromoprotein is expressed in presence of oxygen. | The mold was not hermetically closed, the chromoprotein is expressed in presence of oxygen. | ||
| - | [[File:Paris_Saclay_Space_Cake_1.jpeg| | + | [[File:Paris_Saclay_Space_Cake_1.jpeg|600px|center]] |
It looks like blue tasty space cake but... | It looks like blue tasty space cake but... | ||
| - | [[File:Paris_Saclay_Space_Cake_2.jpeg| | + | [[File:Paris_Saclay_Space_Cake_2.jpeg|600px|center]] |
...when unmolded, you can only see a little ring of blue at 100% ( non-diluted preculture ).In fact, bacteria didn't grow. | ...when unmolded, you can only see a little ring of blue at 100% ( non-diluted preculture ).In fact, bacteria didn't grow. | ||
| - | + | There are no nutrients in agar, bacteria won't grow without appropriate medium. | |
I can't use LB because of its color. I must find a colorless medium. | I can't use LB because of its color. I must find a colorless medium. | ||
| + | |||
| + | ==Photo of the Day== | ||
| + | [[File:Paris Saclay 13_august.jpg|600px|center]] | ||
| + | |||
| + | '''Members present:''' | ||
| + | * Instructors and advisors: Alice, Solenne and Sylvie. | ||
| + | * Students: Eugène, Fabio, Hoang Vu, Mélanie, Romain Sean and Terry. | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 14:51, 14 October 2014
Contents |
Wednesday 13th August
Lab work
Lemon scent
culture of pJBEI6409 bacteria
by Mélanie
culture in LB of some clones obtain yesterday
Gel electrophoresis of BBa_K762100
by Sean
Samples used: 5µl from each tube prepared 12th August
Contents of the two tubes yielding the highest intensity were pooled.
PCR clean-up of BBa_K762100
by Sean
PCR clean-up was conducted on the contents of the two tubes pooled earlier.
Gel electrophoresis of BBa_K762100
by Sean
Sample used: the result of the PCR clean-up conducted today.
Concentration of BBa_K762100 is very low — the band is only slightly visible.
Cloning BBa_K517003 and p cola in pGEMTeasy
by Eugene and Sean
| components | volumes |
|---|---|
| PCR purified p cola/ BBa_K517003 | 7 μl |
| DreamTaq buffer 10X | 1 μl |
| dATP | 2 µl |
| DreamTaq | 1 µl |
incubate for 30 minutes at 70°C
| components | volumes |
|---|---|
| 2xT4ligase buffer | 10 μl |
| pGEMTeasy | 1 μl |
| DNA insert | 4.7 µl at 16.8 ng/µl |
| 2T4DNA ligase | 1 µl |
| H2O | 3.3 µl |
| components | volumes |
|---|---|
| 2xT4ligase buffer | 10 μl |
| pGEMTeasy | 1 μl |
| DNA insert | 5.6 µl at 16.8 ng/µl |
| 2T4DNA ligase | 1 µl |
| H2O | 2.4 µl |
Culture of transformed bacteria
We used 3 colony of bacteria transformed with PJBEI+Apra (11/08) to lauch 3 cultures
-two cultures in 5mL LB+Apra (1/2000)
-one culture in 20mL LB+Apra (1/2000)
37°C - 24h
by Laetitia
Human Practices
Art & Design
by Terry
Unmolding yesterday's preparation. The mold was not hermetically closed, the chromoprotein is expressed in presence of oxygen.
It looks like blue tasty space cake but...
...when unmolded, you can only see a little ring of blue at 100% ( non-diluted preculture ).In fact, bacteria didn't grow.
There are no nutrients in agar, bacteria won't grow without appropriate medium. I can't use LB because of its color. I must find a colorless medium.
Photo of the Day
Members present:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Eugène, Fabio, Hoang Vu, Mélanie, Romain Sean and Terry.
