Team:Evry/Notebook/Sensing/PCBs/08-21-2014

Sensor construction hbpR/PhbpC and hbpR2/PhbpR1
The part BBa_J23114 was located on 2014 Distribution kit plates and resuspended with 10 µL sterile water. That permits to obtain a DNA concentration around 0.2 ng/µl (according to the registry. Solution was transferred into 1 ml eppendorf tubes and stored at -20°C.
Transformations of registry parts (BBa_B0015, E1010 and J23114) and vector PSB1C3G3 were performed to obtain colonies with plasmid containing parts for future amplifications.
The transformation was performed on DH5 alpha ''E. coli'', as followed:

1. Remove E. coli competent tubes from -80°C and keep it on ice
2. Add 3 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently
3. Incubate 10 minutes on ice
4. Perform an heat shock 30 seconds at 42°C
5. Incubate 2 minutes on ice
6. Add 2 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm
7. Plate 200 µl of PSB1C3G3, BBa_B0015 and E1010 on a chloramphenicol LB agar plate or ampiciline LB agar plate for BBa_J23114
8. Incubate plate overnight at 37°C

Table 4: Received primers with numbers and sequences for PCB sensing constructions