IGEM:IMPERIAL/2007/Projects/Experimental Design: Difference between revisions
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*Several Problems have been identified, we have begun to address them. [[IGEM:IMPERIAL/2007/Projects/Experimental Design/Problems| Link here to problems.]] | *Several Problems have been identified, we have begun to address them. [[IGEM:IMPERIAL/2007/Projects/Experimental Design/Problems| Link here to problems.]] | ||
*We are currently trying to improve our methodology to get better results. [[IGEM:IMPERIAL/2007/Projects/Experimental Design/Improve Methodology| Link to improving our Methodology]] | *We are currently trying to improve our methodology to get better results. [[IGEM:IMPERIAL/2007/Projects/Experimental Design/Improve Methodology| Link to improving our Methodology]] | ||
*<font color=red> Problems with dsRED, it has failed quality control</font> | |||
*<font color=red> dsRED | *Currently trying to purify LuxR and GFP. | ||
==Time table== | ==Time table== | ||
| Line 85: | Line 85: | ||
|- style="background:#d4cae2" | |- style="background:#d4cae2" | ||
| style="background:#aa99c5" | '''Wednesday''' | | style="background:#aa99c5" | '''Wednesday''' | ||
|| || <br> Purify LuxR if plasmid arrives || | || || <br> Purify LuxR if plasmid arrives || <font color=red> problems with LuxR pufification<font color=red> | ||
|- style="background:#d4cae2" | |- style="background:#d4cae2" | ||
| style="background:#aa99c5" | '''Thursday''' | | style="background:#aa99c5" | '''Thursday''' | ||
| Line 91: | Line 91: | ||
|- style="background:#d4cae2" | |- style="background:#d4cae2" | ||
| style="background:#aa99c5" | '''Friday''' | | style="background:#aa99c5" | '''Friday''' | ||
|| Test construct with 5 AHL concentrations - 25 °C || Begin purification of LuxR || | || Test construct with 5 AHL concentrations - 25 °C || Begin purification of LuxR ||Test Purified LuxR with construct 2?? | ||
|} | |} | ||
<br> | <br> | ||
Latest revision as of 02:35, 21 September 2007
We have identified three phases of experiments for each of the project:
- Phase 1 - Intial testing of the promoters and test constructs in vitro
- Phase 2 - Testing/Validation of the test constructs
- Phase 3 - Testing/Validation of final constructs
In addition any overlap between the experiments of the projects has been identified and considered when making the experimental design.
Status
- Several Problems have been identified, we have begun to address them. Link here to problems.
- We are currently trying to improve our methodology to get better results. Link to improving our Methodology
- Problems with dsRED, it has failed quality control
- Currently trying to purify LuxR and GFP.
Time table
General timetable
| Week 8 | Week 9 | Week 10 | |
| Monday | - | Refine the wiki | Planning the week |
| Tuesday | Prepare GFP solution-calibration Construct Calibration Curve Refine the wiki |
Testing to reduce evaporation | |
| Wednesday | Test Home made cell extract Test varying levels of cell extract |
PROBLEM-Wrong GFP!!!! | 24hr experiment with pTet using paraffin oil |
| Thursday | |||
| Friday | Prepare GFP solution-calibration Construct Calibration Curve |
CBD
| Week 8 | Week 9 | Week 10 | |
| Monday | - | ||
| Tuesday | Phase 1 - test pT7 | ||
| Wednesday | |||
| Thursday | Test 20 and 37 °C, Test GFP degradation at 20 and 37 °C |
24 Hour Exp Room Temp | |
| Friday | Second day testing 20 and 37 °C | Define the thresholds of temperature |
ID
| Week 8 | Week 9 | Week 10 | |
| Monday | - | Purify LuxR | |
| Tuesday | test pLux in-vitro - 1nM AHL | Refine AHL concentrations range 25°C Purify LuxR if plasmid arrives |
Purification of LuxR - problems |
| Wednesday | Purify LuxR if plasmid arrives |
problems with LuxR pufification | |
| Thursday | Clone in plasmid | ||
| Friday | Test construct with 5 AHL concentrations - 25 °C | Begin purification of LuxR | Test Purified LuxR with construct 2?? |
