Difference between revisions of "Template:IONIS Paris/Notebook/09-09-15"

E.coli DH5 alpha transformation

Transformation of competent cells with pSB1C3-GFP and pSB1C3-H/E (ratio 3:1, 1:1, 1:3) with 3 µl of plasmid.

Liquid cultures on plates from 08/09

Measurements of DO595 and DO405 (wtGFP) all the day long

Typical profile of colonies transformed with pDawn-HOKD under light exposition

Continuous bacterial growth despite light exposure not expected

Typical profile of colonies transformed with pDawn-CCDB under light exposition

Interesting profile with a diminution of the DO595 after it reach 0,6

Typical profile of colonies transformed with pDawn-Holin/Endolysin under light exposition

Continuous bacterial growth in spite of light exposure not expected

But some clones get the following profile

Very interesting profile, the DO600 fall down earlier than for all others clones

Typical profile of colonies transformed with pDawn-GFP under light exposition

Not interesting, all curve are proportional such as there were no production of GFP

PCR colony for pSB1C3-pDawn I-V and pSB1C3-VVD YC-VVD YN

Mix PCR preparation:

Tube	pSB1C3-pDawn I-V	pSB1C3-VVD YC-VVD YN
MQ Water	40 µL	40 µL
RB Buffer	5 µL	5 µL
Mg2+	1,5 µL	1,5 µL
dNTP 10 µM	1 µL	1 µL
Primer Fwd	pDawn Fwd 2, 0,5 µL	pSB1C3 Fwd 2, 0,8 µL
Primer Rev	pDawn Rev 2, 0,5 µL	pSB1C3 Rev 2, 0,8 µL
Taq Pol	0,5 µL	0,5 µL

XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX

Liquid culture

pSB1C3-pDawn, colonies 1 to 5 (there were nothing with PCR colony but we have to try…)
pSB1C3-VVD YC-VVD YN, colony 1

9 September 15