Biobrick

Asaia

• We made pre-culture and plates (on both Kan and Gly alone) of the recovered WT Asaia
• Made PCR of the promotor + RBS and ran gel -> it worked

Immunotoxine

We'll made some GLY stock of E-coli containing the immunotoxine plasmid (shiva). So today we transform E-coli with the plasmid and we plated on Amp overnight.

Biobrick

We try to begin to construc two new biobrick today but we made mistake. We'll start again tomorrow.

Immunotoxine

We've pick up some colony of transformed cells from yesterday and make liquid culutre of them. Tomorrow we'll make some GLY stock and a gel to control. With this stock we can begin to work safely with the immunotoxine.

Asaia

To test our asaia origin sequence, we take a BV with (1) asaia origin (2) Amp resistance (3) death casette and we will throw away the death casette and just transform asaia with this simpel plasmid. Today we just make a liquid culture from a GLY stock with E-coli containing this plasmid.

Biobricks

We begin to construc a new Biobrick containing (1) Base Vector (2) Strong or Weak promotor (3) RBS. Today we made digestion, ligation and E-coli transformation. Plated overnight on Amp.

Drosophile

Persistence of bacteria in drosophilia's gut. We infected (fed) flies with Asaia. We also infected flies with persistant or pathogenic bacteria (Pseudomonas in control experiment). Then we crushed, diluted and plated infected flies after 3h hours from infection. We will plate flies after 24 and 48 hours as well and see how many colonies appear on the plates. We hope to get an idea of the persistency of Asaia in the gut of Drosophila.

Drosophila

• Same thing as yesterday afternoon, Making plates to get the CFU-count
• Plates of yesterday with Pseudomonas look promising, the cells grew and the dilution was there.

Biobrick

Digestion of Immunotoxine Other jobs failed and we discovered we have to restart many digestion/ligation Digestion C3+Promotor(weak/strong)+RBS Digestion C3+Promotor(w/s)+RBS+Immuno

Drosophilae

• Continued yesterday's experiment with plating, this time the 48h persistence.
• Unfortunately the plates from yesterday don't look good. We might have killed all the bacteria by washing the flies in ethanol for too long.

Asaia

• Prepared a wild type recovery pre-culture to obtain competent cells. We're repeating the procedure from two weeks ago, because what we obtained then didn't seem to be the wild type (it grew on plates with Kanamycin).

Biobrick

Digestion of Immunotoxine Other jobs failed and we discovered we have to restart many digestion/ligation Digestion C3+Promotor(weak/strong)+RBS Digestion C3+Promotor(w/s)+RBS+Immuno

BioBricks

Today we came to check our culture of E-coli transformed with the plasmid C3 + Promotor + RBS (+ Immunotoxine) but no plate have grown!!

Asaia

We also transformed E-coli 3.1 with the Base Vector (BBa_I51020) + Asaia Origin + Resistance.

Asaia

Today we just come and check our transformed cells of yesterday. We have colonies on two plates. Work continue tomorrow.