Team:Paris Bettencourt/Notebook/Trojan Horse/Tuesday 2nd July.html

Trojan Horse

ASDF

2^nd July

Place your twit here

Place your note here

Aude and Vincent

glycerol stock

from overnight culture of MG1655-6300 O/N : T001

• Centrifuge 4000rpm, 10 minutes,

• take out liquid

• resuspend cells in 1mL glycerol, 2mL LB

• separate in two cryotubes, one for the -80°C, one for the -20°C

Electroporation

- making MG1655-6300 competent (Aude’s protocol for electrocompetent cells & electroporation [ref needed])

- test of the competent cells (negative

-transforming with pCOLADuet-1 : Ec2 =>

-transforming with pACYCDuet-1 : Ec2 =>

- plating 3 different quantity of cells (20ul, 50uL, 100uL) respectively on Cm and Kan plates (dilution 1000x for the antibiotics)

03/07

Aude and Vincent

Transfo results :

• negative controls are negative.

• Transformed cells grew on plates

• in the evening launch of 5mL Lb cultures from clones on the plates

Igem Buffer for chemical competent cells

Igem protocol for chemical competent cells

Launch overnight culture of NEB turbo

5mL LB inoculated with a clone from plate of drug team