Template:IONIS Paris/Notebook/16-09-15
Miniprep
Liquid culture (pSB1C3-Holin/Endolysin x4 and pSB1C3-pDawn x2)
Liquid culture from the bacterial growth kinetic experiment (pDawn-Holin/Endolysin x2)
Digestion
| Tube | Miniprep | pDawn-H/E |
|---|---|---|
| H2O MQ | 2 μL | 3 μL |
| Buffer 2.1 | 1 μL | 1 μL |
| DNA | 5 μL | 5 μL |
| EcoRI | 1 μL | 1 μL |
| PstI | 1 μL | - |
37°C, 1h
Expected results
Results
There were no expected profile of digestion excepted for the first and the third wells with pDawn-Holin/Endolysin.
Digestion
| Tube | Biobrick |
|---|---|
| H2O MQ | 5 μL |
| Buffer 2.1 | 1 μL |
| DNA | 3 μL |
| EcoRI | 1 μL |
37°C, 1h
Liquid culture of bacteria transformed with pSB1C3-nanoluc (x5)
Liquid culture on 96 wells plate
When 0,6 < OD600 < 0,8, induction with arabinose or IPTG
Incubate 5h at 30°C; for BBa_K325909, measure the kinetic during the 5h of induction and take the maximum value
Add 50µL of NanoGlow substrate (Promega) for 50 uL of bacteria
Measure kinetic over 120 min and take maximum value for culture with BBa_K1159001 and T7-nanoluc
16 Sept. 15