Team:Paris Saclay/Notebook/August/13

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==Lab work==
==Lab work==
-
''by Melanie''
+
===Lemon scent===
 +
====culture of pJBEI6409 bacteria====
 +
''by Mélanie''
-
Preparation of DH5a CaCl2 Competent cell
+
culture in LB of some clones obtain yesterday
 +
 
 +
====Gel electrophoresis of BBa_K762100====
 +
''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.
-
===D - Lemon scent===
 
====Gel electrophoresis of BBa_K762100====
====Gel electrophoresis of BBa_K762100====
''by Sean''
''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====
-
''by Eugène and Sean''
+
''by Eugene and Sean''
 +
 
 +
{| class="wikitable centre" width="50%"
 +
|+ P cola and BBa_K517003 cloning
 +
|-
 +
! scope=col | components
 +
! scope=col | 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
 +
 
 +
{| class="wikitable centre" width="50%"
 +
|+ Geraniol synthase
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|2xT4ligase buffer
 +
|10 μl
 +
|-
 +
|pGEMTeasy
 +
|1 μl
 +
|-
 +
|DNA insert
 +
|4.7 µl at 16.8 ng/µl
 +
|-
 +
|2T4DNA ligase
 +
|1 µl
 +
|-
 +
|H<sub>2</sub>O
 +
|3.3 µl
 +
|}
 +
 
 +
{| class="wikitable centre" width="50%"
 +
|+ Pinene synthase
 +
|-
 +
! scope=col | components
 +
! scope=col | volumes
 +
|-
 +
|2xT4ligase buffer
 +
|10 μl
 +
|-
 +
|pGEMTeasy
 +
|1 μl
 +
|-
 +
|DNA insert
 +
|5.6 µl at 16.8 ng/µl
 +
|-
 +
|2T4DNA ligase
 +
|1 µl
 +
|-
 +
|H<sub>2</sub>O
 +
|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 [https://2014.igem.org/wiki/index.php?title=Team:Paris_Saclay/Notebook/August/12 yesterday]'s preparation.
 +
The mold was not hermetically closed, the chromoprotein is expressed in presence of oxygen.
 +
 
 +
[[File:Paris_Saclay_Space_Cake_1.jpeg|600px|center]]
 +
 
 +
It looks like blue tasty space cake but...
 +
 
 +
[[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.
 +
 
 +
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==
 +
[[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

P cola and BBa_K517003 cloning
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

Geraniol synthase
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
Pinene synthase
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.

Paris Saclay Space Cake 1.jpeg

It looks like blue tasty space cake but...

Paris Saclay Space Cake 2.jpeg

...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

Paris Saclay 13 august.jpg

Members present:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Eugène, Fabio, Hoang Vu, Mélanie, Romain Sean and Terry.

Back to the calendar

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