Team:Paris Bettencourt/Notebook/Trojan Horse/Friday 30th August.html


Trojan Horse

30th August

Place your twit here

attempt to silence LacZ on MG1655, F+ with litmusGFP-LacZ1 & litmusGFP-LacZ2


Silencing experiments (LacZ)

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

-diluting 100x O/N of MGZ1, F+ -pCola in LB-Kan

- At OD 0,7 : 5 tubes. Put 1 mL of cells in each tube. Add in tubes respectively : nothing, 100uL phages of litmusGFP-LacZ1 & litmusGFP-LacZ2, litmusGFP-Cm1 & litmusGFP-Cm2

litmusGFP-Cm1 & litmusGFP-Cm2 are used as negative control.

- Incubate 45 minutes at 37°C

- Serial Dilute every tube until 10-5. Plate for each tube 100ul of 10-5 and 10-3 dilutions on IPTG+Xgal and LB and 100uL on AMP+IPTG+Xgal and LB.