From Monday the 18th of July to Friday the 22nd of July 2011

Monday

---

Concerned parts : RBS-GFP-LVA (Cn),BBa_J23119 ( 18A, plate 1 )(Amp), Curli (Cn), ompR234 (Cn)

Start of 5mL liquid cultures from a Petri dish culture:
5mL sterile LB
50µL antibiotic
bacteria
Incubation at 37°C

Later, start of 150mL liquid cultures from the previous 5mL cultures
150 mL sterile LB
300 µL from the grown 5mL bacterial culture
1.5 mL antibiotic ( Amp or Cn )
Incubation at 37°C overnight.


10X concentration and plating of 100 µL of transformed bacteria from the previous week : 2M+24E ( amp ), Curli + GFP ( Kan ), Kan + 2L ( Kan ), positive control ( Puc18 plasmid ), negative control ( water )
Incubation at 37°C

Tuesday

---

Midiprep to extract DNA from the previous liquid cultures.
Nanodrop quantification :
OmpR234 : 59.5 ng/µL
GFP : 109.5 ng/µL
18A : 73.7 ng/µL
Curli : 82.6 ng/µL
The 260/280 ratio is lower than 2 in all cases.

Wednesday

---

Digestion of several biobricks : RBS, GFP, YFP, Curli, OmpR234. Electrophoresis to control the digestions.

Thursday

---

Standard or 3A ligation of the previously cut biobricks.TSS transformation into NM522.

Friday

---

Start of 5mL liquid cultures of selected individual colonies. Extraction of the plasmids to test the presence of the insert.