Team:Paris Saclay/Notebook/September/4

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

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 (Topo plasmid)

First I had to add AAA to the PCR

72° 15'

and

30' at room temperature

Transformation

Transformation with competent bacteria

add DNA (Top vector our pPSII)

30' on ice 45' 42°c 30' at 37°

spread on petri dish