IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 1.1: Difference between revisions
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* [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 '''pT7-GFP'''] testing failed because it was not properly induced. It will be re-tested on the 17/08/2007 after it has been induced with IPTG and left overnight to allow for a measurable GFP amount to be produced. IPTG is a T7 inducer for the E.Coli strain used (BL21). | * [http://parts.mit.edu/registry/index.php/Part:BBa_E7104 '''pT7-GFP'''] testing failed because it was not properly induced. It will be re-tested on the 17/08/2007 after it has been induced with IPTG and left overnight to allow for a measurable GFP amount to be produced. IPTG is a T7 inducer for the E.Coli strain used (BL21). | ||
20/8/2007 | |||
pT7 was tested the next day and no fluorescence was observed. Two reasons were hypothesized: The bacteria cells were in stationary phase; or that the pT7 construct is not working. | pT7 was tested the next day and no fluorescence was observed. Two reasons were hypothesized: The bacteria cells were in stationary phase; or that the pT7 construct is not working. | ||
21/8/2007 | |||
pT7 was found to not work in vivo. Its fluorescence reading was negligibly higher than a normal cell culture, and hence our conclusion is that the gene construct is faulty. (This was tested by comparing the plasmid length after a restriction enzyme digest, and the bands did not tally with the theoretical version of the plasmid construct.) | |||
* [http://parts.mit.edu/registry/index.php/Part:BBa_I719022 '''pCI-GFP'''] testing has been postponed due to delays in the making of the construct. Testing will be carried out as soon as the construct is ready. | * [http://parts.mit.edu/registry/index.php/Part:BBa_I719022 '''pCI-GFP'''] testing has been postponed due to delays in the making of the construct. Testing will be carried out as soon as the construct is ready. | ||
Revision as of 08:31, 21 August 2007
Results summary
- pTet-GFP has been successfully tested in vivo. The levels of fluorescence of the construct where comparable to the levels of our positive control (200x dilution of GFP) meaning that GFP was actually produced. Therefore pTet-GFP works in E.Coli.
- pT7-GFP testing failed because it was not properly induced. It will be re-tested on the 17/08/2007 after it has been induced with IPTG and left overnight to allow for a measurable GFP amount to be produced. IPTG is a T7 inducer for the E.Coli strain used (BL21).
20/8/2007 pT7 was tested the next day and no fluorescence was observed. Two reasons were hypothesized: The bacteria cells were in stationary phase; or that the pT7 construct is not working. 21/8/2007 pT7 was found to not work in vivo. Its fluorescence reading was negligibly higher than a normal cell culture, and hence our conclusion is that the gene construct is faulty. (This was tested by comparing the plasmid length after a restriction enzyme digest, and the bands did not tally with the theoretical version of the plasmid construct.)
- pCI-GFP testing has been postponed due to delays in the making of the construct. Testing will be carried out as soon as the construct is ready.
- A 1 hour run (taking measurements every 2.5 min) was also conducted to test the variation on the expression levels of pTet-GFP over time.
| Complete set of results and raw data . |
