Week 14
From 2007.igem.org
(Difference between revisions)
| Line 11: | Line 11: | ||
::-pLac-GFP+I76303; | ::-pLac-GFP+I76303; | ||
::-I763012+I763020(I763032). | ::-I763012+I763020(I763032). | ||
| + | |||
| Line 22: | Line 23: | ||
*Run on electrophoresis gel. | *Run on electrophoresis gel. | ||
*I763031, I763036 and pSB4A3 band extraction. | *I763031, I763036 and pSB4A3 band extraction. | ||
| + | *Transformation of | ||
| + | ::-pTet-RFP-Plac GFP; | ||
| + | ::-I763031; | ||
| + | ::-I763032. | ||
| + | *Ligation for | ||
| + | ::-I763031+pSB4A3; | ||
| + | ::-I763032+pSB4A3. | ||
'''Experimental Setup''' | '''Experimental Setup''' | ||
| Line 34: | Line 42: | ||
::'''10/04/07''' | ::'''10/04/07''' | ||
| + | *Trasformation of | ||
| + | ::-I763031+pSB4A3; | ||
| + | ::-I763032+pSB4A3. | ||
| + | *We inoculate I763034,I763031+pSB4A3 and I763032+pSB4A3 (in 6 ml). | ||
| Line 39: | Line 51: | ||
::'''10/05/07''' | ::'''10/05/07''' | ||
| - | + | *Stock preparation (500 ul) of I763034,I763031+pSB4A3 and I763032+pSB4A3. | |
| + | *Miniprep of I763034,I763031+pSB4A3 and I763032+pSB4A3. | ||
| + | *Control digestion of | ||
| + | ::-I763034 with Xba/Pst1; | ||
| + | ::-I763031 with Eco/Xba and Xba/Pst1; | ||
| + | ::-I763032 with Eco/Xba and Xba/Pst1. | ||
| + | *Run on electrophoresis gel and I763032(Eco/Xba and Xba/Pst1) band extraction. | ||
[[Bologna | Back]] | [[Bologna | Back]] | ||
Revision as of 09:33, 8 October 2007
- 10/01/07
- Meeting: Ethical issue.
- 10/02/07
- Digestion of PSB4A3;
- Run on electrophoresis gel.
- pSB4A3 and Ptet-RFP band extraction.
- Ligation for
- -pLac-GFP+I76303;
- -I763012+I763020(I763032).
- 10/03/07
- Digestion of:
- -pLac-Lacy-GFP with Eco/Pst1;
- -I763031 with Eco/Pst1;
- -I763036 with Eco/Pst1;
- -pSB4A3 with Eco/Pst1.
- Run on electrophoresis gel.
- I763031, I763036 and pSB4A3 band extraction.
- Transformation of
- -pTet-RFP-Plac GFP;
- -I763031;
- -I763032.
- Ligation for
- -I763031+pSB4A3;
- -I763032+pSB4A3.
Experimental Setup
- To have an higher precision for our measurements, we have decided to get, for the same field of view on the same sample of bacteria solution, either a short film segment (from which we can extract frames) and a Microsoft Excel file, which contains a ten seconds interval of values from the photomultiplier output, taken at a 100Hz frequency.
- In order to have the same focus for the microscope's oculars, the photocamera and the photomultiplier we have to calibrate our system by changing the position of both camera and photomultiplier.
- Finally, we check that the photomultiplier field of view is the same size as the microscope's one: by moving the appropiate diaphragms, we adjust the previously done settings.
- 10/04/07
- Trasformation of
- -I763031+pSB4A3;
- -I763032+pSB4A3.
- We inoculate I763034,I763031+pSB4A3 and I763032+pSB4A3 (in 6 ml).
- 10/05/07
- Stock preparation (500 ul) of I763034,I763031+pSB4A3 and I763032+pSB4A3.
- Miniprep of I763034,I763031+pSB4A3 and I763032+pSB4A3.
- Control digestion of
- -I763034 with Xba/Pst1;
- -I763031 with Eco/Xba and Xba/Pst1;
- -I763032 with Eco/Xba and Xba/Pst1.
- Run on electrophoresis gel and I763032(Eco/Xba and Xba/Pst1) band extraction.
