Team:Paris Bettencourt/Notebook/Trojan Horse/Wednesday 4th September.html

From 2013.igem.org

Trojan Horse

4th September


Results of silencing experiments

It works !

Miller assay

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Redo the silencing experiment

- Launch O/N culture of producing phages cells ; centrifuge ; take supernatant and filter it.

-diluting 100x O/N of MGZ1, F+ -pACYc in LB-Cm (25 ug/ul)

- At OD 0,7 : 5 tubes. Put 900 uL of cells in each tube. Add in tubes respectively : nothing, 100uL phages of litmusGFP-LacZ1, litmusGFP-Cm1 & litmusGFP-Cm2

litmusGFP-lacZ1 & litmusGFP-lacZ2 are used as negative control.

- Incubate 45 minutes at 37°C

- Serial Dilute every tube until 10-5. Plate 100ul for each tube according to this dilution table

LB

Cm 25 ug/ul

Cm 125 ug/ul

Cm 250ug/ul

Cm 500 ug/ul

non infected

10^-4

10^-4

10^-4

10^-4

10^-4

sRNA anti lacz

10^-4

10^-4

10^-3

10^-3

10^-3

sRNA Cm1

10^-4

10^-4

10^-3

10^-3

10^-3

sRNA Cm2

10^-4

10^-4

10^-3

10^-3

10^-3

sRNA Cm3

10^-4

10^-4

10^-3

10^-3

10^-3

sRNA Cm4

10^-4

10^-4

10^-3

10^-3

10^-3