Team:Paris Saclay/Notebook/September/4

From 2014.igem.org

(Difference between revisions)
(PCR verification)
(Verfication of the pPS3/4/5 plasmid)
Line 8: Line 8:
I use the same PCR condition than [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase September 2] but with the right primer and plasmid
I use the same PCR condition than [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2#Limonene_synthase September 2] but with the right primer and plasmid
-
[photo]
+
[[File:0309 Vérif PCR GS CAD PS LS Chromo.jpg|400px]]
We can see that we don't have any insert in our plasmid
We can see that we don't have any insert in our plasmid

Revision as of 12:06, 7 September 2014

Contents

Lab Work

D- Lemon Scent

By Mélanie

Verfication of the pPS3/4/5 plasmid

Due to the strange results obtained last day, I do a PCR of the insert with the differents plasmid : I use the same PCR condition than September 2 but with the right primer and plasmid

0309 Vérif PCR GS CAD PS LS Chromo.jpg

We can see that we don't have any insert in our plasmid

PCR verification

We find some other PCR of BBa K762100 and GS and BBa_K517003 so we check it by electrophoresis

0309 Vérif PCR GS CAD PS LS Chromo.jpg

And I purify the PCR

0309 PCR cleanup GS PS CAD.jpg


Well 1 = Ladder Well 2 = GS Well 3 = PS Well 4 = CAD

We can see that we don't have lost a lot of our PCR product

cloning

Cloning of PCR purification in a TA plasmid First I had to add AAA to the PCR

component volume
buffer 1μl
dATP 1μl
PCR 7.5μl
Taq 0.5μl

72° 15'

and

component volume
H2O 1μl
buffer 1μl
vector 1μl
cloning 1μl

30' at room temperature

and transformation with competent bacteria add DNA 30' on ice 45' 42°c 30' at 37°

spread on petri dish