Team:Paris Saclay/Notebook/September/8

From 2014.igem.org

(Difference between revisions)
m (Monday 8th September)
m (Photo of the Day)
Line 144: Line 144:
==Photo of the Day==
==Photo of the Day==
-
[[File:Paris Saclay 8_september.jpg|600px|center]]
+
[[File:Paris Saclay 8_september.jpg|150px|center]]
 +
 
 +
[http://iramis.cea.fr/llb/Phocea/Pisp/visu.php?id=35&uid=%20alexei.grinbaum Alexei Grinbaum] , one of those few we have interviewed during this month of september.
 +
 
 +
To learn more about the ethic aspect of our project, just click [http://2014.igem.org/Team:Paris_Saclay/Ethics here]
 +
 
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 00:36, 14 October 2014

Contents

Monday 8th September

Lab Work

D-Lemon Scent

PCR with bacteria

From the petri dish (made here, I select some clone to do a PCR : clones 1-2-3 for cad clones 4-5 for PS clones 6 for LS

component volume
H2O 41.5μl
buffer 5μl
dNTPs 1μl
Primer 1 1μl
Primer 2 1μl
bacteria (about 1µl = 1 colony)
Dream taq 0.5μl

Primer used:

For cloning in Topo vector = Pu Pr (universal primer)

For pPS2 = iPS 66/67

Cycle step Temperature Time Cycle

Bacteria lysis

95°C

5 min

1

Denaturation 94°C 30 s 25
Annealing 50°C 25 s 25
Extension 72°C 1 min 25
Final extension 72°C 10 min 1
Final extension 8°C hold 1

0809 PCR clones.jpg

We identify that we have success with to clone of PS. (well 5-6)

Liquide culture of clones

The PS clone that reveal to be ok are cultivate in LB + AMP médium

Checking PCR

Electrophoresis of the PCR made Friday

LS PS GS CAD (and chromo) (2 enzymes = vent taq and dream taq)

0809 PCR LSPSGSCADCRHOMO vent dream.jpg

Cloning

With this PCR, I do a cloning in a Topo Cloning vector (Zero Blunt TOPO PCR Cloning Kit) (LS GS and CAD)

component volume
PCR product 1μl
Salt 1μl
Vector 1 μl
H2O 3μl


Between 5 and 30 min at room temperature

Transformation

Add 50µL of bacteria to the clonage previously done

30min on ice 45 sec at 42°C

add 1ml of LB

1hour at 37°

Spread on a petri dish

B-Contruction of the fusion protein

We check the sequence of the fusion protein and identify a clone with our chromoprotein = W6

Photo of the Day

Paris Saclay 8 september.jpg

Alexei Grinbaum , one of those few we have interviewed during this month of september.

To learn more about the ethic aspect of our project, just click here


Back to the calendar