BioBricks

The transformation of E-coli with the circular C3 didn't work.... The big problem is that we haven't no more C3 in stock. So to recover some C3, we decide to transform E-coli DH5alpha with the vector C3 + RFP.

We made a colony PCR with the E-coli transformed with C3 + [Strong promotor | Weak promotor] + [ nothing | Immunotoxine]. The C3 + W/S work but the immunotoxine didn't. => liquid culture from the colony how work.

Asaia

Drosophile

BioBricks

We do some GLY stock from the colony how work yesterday. Then with the rest of the liquid culture we made miniprep to send the plasmid to sequencing. We also do some liquid culutre with E-coli transformed with C3 + RFP. We'll do GLY stock tomorrow. We made a colony PCR of E-coli transformed with C3 + Asaia origin but the result fail. To try to construct C3 + [Strong promotor | Weak promotor] + [Immunotoxine | P25 | P28]. Two step today : digestion of all plasmids and overnight ligation. We send 4 plasmid to sequence :

1. C3 + Weak promotor
2. C3 + Strong promotor (colony #7)
3. C3 + Strong promotor (colony #8)
4. C3 + Asaia origin + Kan
5. C3 + Asaia origin + Amp

Asaia

Result from yesterday seem to indicate that Asaia Wild Type is naturally resistant to Amp and Ch. We redo some test to be sure we've do any mistake.

Drosophile

Biobricks

The plasmids we sent to sequencing came back positive. We have now 4 biobricks.

1. C3 + Asaia_origin + Kan
2. C3 + Asaia_origin + Amp
3. C3 + weak promoter + RBS
4. C3 + strong promoter + RBS

We also ran a gel with C3 + asaia origin. It looks good.

Asaia

Drosophilae

We restarted the experiment with the drosophila. This time, we crushed 15 flies and we washed them in ethanol for only 2 seconds (maximum). We also did the plating of the flies 3h after infection

Biobricks

We did a digestion and ligation of our 2 vectors C3 + S and C3 + W with the immunotoxin, P25 and P26. In the end, we should have 6 new biobricks.

Asaia

We are still not sure if Asaia "WT" is resistant to kanamycin or not. Maybe we will have to use another resistance in our biobricks, TetR for instance.

Drosophilae

We are doing the crushing and plating of the drosophilae that were infected 24h ago.