Team:UNIPV-Pavia/Notebook/Week17

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Notebook

Week 1	Week 2	Week 3	Week 4	Week 5	Week 6	Week 7
Week 8	Week 9	Week 10	Week 11	Week 12	Week 13	Week 14
Week 15	Week 16	Week 17	Week 18	Week 19	Week 20	Week 21

09/8/08

We infected 9 ml of LB + Amp with 30 µl of Lig.30(3), Lig.31, Lig.22, Lig.13 and two falcon tubes for Lig.a.

We received 3OC6HSL from Sigma! we resuspended it in ddH2O, we prepared some stocks and stored them at -20°C.

09/9/08

Ligations:
- Lig.31-Lig.30 (="Lig.34")
- Lig.31-Lig.22 (="Lig.35")
- Lig.30-Lig.13 (="Lig.T5" for green fluorescence test)

We induced one of the two Lig.a overnight culture with 3OC6HSL 1 µM. We incubated the two cultures for 1 hour and then watched TRITC channel at microscope. (We didn't synchronize the two cultures, but performed a qualitative test for luxR mutated protein integrity evaluation).

Fluorescence test results: HSL induced Lig.a show RFP expression, while Lig.a without HSL showed a weak red fluorescence. Result pictures are not available at the moment.

09/10/08

We transformed/plated ligations. We decided to perform two transformations for each ligation:
- one normal transformation (1 µl of ligation);
- one diluted ligation (1:10)
We decided to try diluted transformations to have less colonies in the plate and to avoid streaking single colonies plates when colonies are not insulated.

09/11/08

Colony PCR for Lig.34, Lig.35 and Lig.T5 (4 colonies for normal transformation plates and 4 colonies for diluted transformation plates).