Team:Paris Saclay/Notebook/September/4

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(PCR verification)
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And I purify the PCR
And I purify the PCR
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Revision as of 12:03, 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

[photo]

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