BioWire Experiments

Experiments

TESTING RECEIVER J06000

Parts: I13272 (AHL receiver with YFP output)

Construction: Transform cells.

Experiments:

Add AHL at different concentrations.
Generate time curves and transfer curves of fluorescence. (use fluorimeter, spectrophotometer)
Two Controls. 1) Same cells in no AHL 2) different cells without construct
Verification by running gel

Questions:

To do Tuesday 6/14:

COA: 1500- If e. coli is ready, mini-prep those suckas. Else, grow them up. mini-prep, retransform,

TESTING SENDER J06001, J06002, I13602(J06003)

Construction: Ligate R0040 and F1610 (LUXI LVA J06001), and R0040 and E0422 (CFP J06003), and R0040 and F1611 (LUXI LVA- J06002). Transform to make sender cell. Also make non-CFP version (?).

Experiments:

Add aTc at different concentrations.
Generate time curves and transfer curves of fluorescence. (use fluorimeter, spectrophotometer)
Two Controls. 1) Same cells in no aTc 2) different cells without construct
Verification by running gel

Questions:

TESTING SENDER AND RECEIVER (in different cells)

Construction:

Receiver: J06000

Experiments:

Add aTc (should know how much to add from previous experiment).
Sender cells should fluoresce CFP, receiver cells should fluoresce YFP.
Controls: Add water instead of aTC, should get no fluorescence (or basal level due to leaky promoters).
Do this experiment on agar. Mix cells (one experiment) and also separate senders and receivers (other experiment--will help us get some idea of distances and diffusion.)

WEISS CIRCUIT TESTING

Parts: J06001, J06002, J06003 (sender cell) and J06004, J06005 (receiver cell)

Construction:

Receiver: J06004 and J06005

Experiments:

TESTING OUR CIRCUIT

Parts: J06007 (LVA LUXI), J06008 (LVA- LUXI), J06004

Construction: J06007 and J06004 OR J06008 and J06004.

Experiments:

Have one sender cell. Add aTc. Compare propagation of signal down line with our cells with propagation of signal down line with Weiss cells (control). We should have stronger propagation.

OTHER CONSTRUCTIONS