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From 2014.igem.org

Contents 1 Lab Work 1.1 D- Lemon Scent 1.1.1 Verfication of the pPS3/4/5 plasmid 1.1.2 PCR verification 1.1.3 cloning

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

And I purify the PCR

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