Team:Paris/25 August 2009

NoteBook
August 25th

 

Lab work
Digestion/Gel purification w/ Soufifi

Digestion of OmpA Linker by SacI and Spe I

Digestion of Ompa Signal by the same enzymes



A41 OmpAsignal PCR OmpAsignal A41 ; 70,4° ; 9s ; matrice:pINK2

Gel purif A41 OmpAsignal A40 TolRII Digestion X/P

Ligation pSB1A3

A41 OmpAsignal ligation topovector

PCR+ check + purification

PCR->A34 (NTER ClyA)

Check gel--> OK

Purification-->ok

Miniprep+Sequencing

Miniprep of all colonie of nter RFP and Cter RFP

Colonie DO:

Nter RFP: for colonie

-1:1,80µg/ml

(Miniprep: 3,33µL+ oligos VF2(1/50): 2,45µL+ H20: 9,22µL)

(Miniprep: 3,33µL+ oligos VR(1/50): 2,45µL+ H20: 9,22µL)

-2:1,90µg/ml

(Miniprep: 3,16µL+ oligos VF2(1/50): 2,45µL+ H20: 9,39µL)

(Miniprep: 3,16µL+ oligos VR(1/50): 2,45µL+ H20: 9,39µL)

-3:1,65µg/ml

(Miniprep: 3,64µL+ oligos VF2(1/50): 2,45µL+ H20: 8,81µL)

(Miniprep: 3,64µL+ oligos VR(1/50): 2,45µL+ H20: 8,81µL)

-4:1,66µg/ml

(Miniprep: 3,61µL+ oligos VF2(1/50): 2,45µL+ H20: 8,94µL)

(Miniprep: 3,61µL+ oligos VR(1/50): 2,45µL+ H20: 8,94µL)

Cter RFP: for colonie

-1:1,80µg/ml

-2:1,90µg/ml

-3:1,70µg/ml

-4:1,60µg/ml