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Team:Paris Saclay/Notebook/August/13
From 2014.igem.org
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Sample used: the result of the PCR clean-up conducted today. | Sample used: the result of the PCR clean-up conducted today. | ||
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====Cloning BBa_K517003 and p cola in pGEMTeasy==== | ====Cloning BBa_K517003 and p cola in pGEMTeasy==== | ||
Revision as of 15:13, 20 August 2014
Contents |
Wednesday 13th August
Lab work
D - 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 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.
I forgot that there are no nutrients in agar alone, bacteria won't grow without appropriate medium. I can't use LB because of its color. I must find a colorless medium.
