Team:Paris Saclay/Notebook/September/3

LabWork

B- Construction of the fusion protein

We made a classic exttraction of plasmids from the liquid cultures.

Check by electrophoresis if it worked and send it for sequencing.

D- Lemon scent

by Mélanie

PCR LS

Electrophoresis of the PCR made Yesterday

well 2-3 = PCR with Dream taq or Vent taq

(well 4-5 = checking of the pps5 Extraction)

The PCR have success so I use the PCR clean up kit to purify it

Electrophoresis after purification

Digestion

In order to clone LS in pPSI, i Digest it by PacI

Ligation

We already have some pPSI digested and dephosphorelated

So I do a ligation:

2 hours at room temperature and over night at 4°

pPS5

Plasmid exctraction using the kit (picture is here)

digestion by SalI

Electrophoresis

We saw that we have something strang with our plasmid but we wil check it tomorrow with a good ladder (the ladder here was normally on the well 1)

pPS3 and pPS4

We digest the plasmid by HindIII to direct our insert

well 1 = ladder well 2-3-4 = pPS3 Well 5-6-7 = pPS4

results are very strange