Team:Paris Saclay/Notebook/September/4


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(Lab Work)
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==Lab Work==
==Lab Work==
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spread on petri dish
spread on petri dish

Revision as of 19:39, 29 September 2014


Lab Work

By Mélanie

D- Lemon Scent

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 of PCR purification in a TA plasmid (Topo plasmid)

First I had to add AAA to the PCR

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

72° 15'


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

30' at room temperature


Transformation with competent bacteria

add DNA (Top vector our pPSII)

30' on ice 45' 42°c 30' at 37°

spread on petri dish

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