Team:Paris Saclay/Notebook/September/2

From 2014.igem.org

(Difference between revisions)
(Created page with "==B-Contruction of the fusion protein== We can see that the blue colonies of E.coli are not blue in a simple petri dish with ampicillin. It's a fail. So, to figure our problem w...")
(→B-Contruction of the fusion protein)
Line 1: Line 1:
-
==B-Contruction of the fusion protein==
+
==Labwork==
 +
 
 +
===B-Contruction of the fusion protein===
 +
 
 +
'' By Hv''
We can see that the blue colonies of E.coli are not blue in a simple petri dish with ampicillin. It's a fail.
We can see that the blue colonies of E.coli are not blue in a simple petri dish with ampicillin. It's a fail.
Line 15: Line 19:
We first did a liquid culture in 5 mL of LB.
We first did a liquid culture in 5 mL of LB.
 +
 +
===D- Lemon scent===
 +
 +
''By Melanie''
 +
 +
Limonene synthase
 +
Migration of the PCR done [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/1 yesterday]
 +
 +
photo
 +
 +
 +
well 1= ladder
 +
well 2-6 = pPS5
 +
other well = LS PCR
 +
As we see, there are nothing on our gel so we conclude that we failed to clone LS in pGMETeasy
 +
 +
pPPS5
 +
 +
the photo in the same as previously: We saw a band but we will check another time by PCR to be sure.
 +
 +
pPS3 and pPS4
 +
 +
Plasmid extraction using the plasmid DNA extraction kit and electrophoresis:
 +
 +
[photo]
 +
 +
We conclude that we have done a good manipulation.
 +
 +
 +
Limonene synthase
 +
As we saw that we don't have anything for LS, We try to do other PCR from the Biobrick (BBA762100). We use two enzyme: Dream taq and Vent taq

Revision as of 18:23, 3 September 2014

Labwork

B-Contruction of the fusion protein

By Hv

We can see that the blue colonies of E.coli are not blue in a simple petri dish with ampicillin. It's a fail. So, to figure our problem we decided to do a sequencing of our plasmids.

We chose 6 colonies from an Xgal-IPTG petri dish:

- 1 completely blue

- 2 blue with a white ring around the blue

- 3 white colony

PS: we found that there is 2 types of blue colonies: completely blue and blue with a white ring around the blue)

We first did a liquid culture in 5 mL of LB.

D- Lemon scent

By Melanie

Limonene synthase Migration of the PCR done yesterday

photo


well 1= ladder well 2-6 = pPS5 other well = LS PCR As we see, there are nothing on our gel so we conclude that we failed to clone LS in pGMETeasy

pPPS5

the photo in the same as previously: We saw a band but we will check another time by PCR to be sure.

pPS3 and pPS4

Plasmid extraction using the plasmid DNA extraction kit and electrophoresis:

[photo]

We conclude that we have done a good manipulation.


Limonene synthase As we saw that we don't have anything for LS, We try to do other PCR from the Biobrick (BBA762100). We use two enzyme: Dream taq and Vent taq

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/September/2"