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Team:Paris Bettencourt/Notebook/Trojan Horse/Tuesday 3rd September.html
From 2013.igem.org
Trojan Horse
ASDF3rd September
Silencing experiments (Cm)
- 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-LacZ2, 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 250ug/ul |
Cm 500 ug/ul |
|
|
non infected |
10^-4 |
10^-4 |
10^-4 |
10^-4 |
|
sRNA anti lacz |
10^-4 |
10^-4 |
10^-3 |
10^-3 |
|
sRNA Cm1 |
10^-4 |
10^-4 |
10^-3 |
10^-3 |
|
sRNA Cm2 |
10^-4 |
10^-4 |
10^-3 |
10^-3 |
|
sRNA Cm3 |
10^-4 |
10^-4 |
10^-3 |
10^-3 |
|
sRNA Cm4 |
10^-4 |
10^-4 |
10^-3 |
10^-3 |
