Difference between revisions of "Part:BBa I13600:Experience"
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| − | The gel analysis and the plasmid miniprep worked quite nicely as expected. But later the construct was further tested and | + | The gel analysis and the plasmid miniprep worked quite nicely as expected. But later the construct was further tested and characterized by combining with the bio-brick BBa_K182005 which contained a tet repressor under the control of a CI operator. So, the CFP expression of BBa_I13600was repressed as expected when tested with the BBa_K182005. With the addition of Anhydrotetracycline (an analogue of tetracycline) the CFP expression was restored and this was confirmed performing the fluorescent microscopy assay. |
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| − | Expression can be | + | Expression can be visualized very nicely under a fluorescent microscope. Even without having ideal filters. We used a GFP filter for the picture below. |
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[[Image:2 I13600 flu60aberdeen2009.jpg|center|400 px]] | [[Image:2 I13600 flu60aberdeen2009.jpg|center|400 px]] | ||
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Since K182005 is on an Ampicillin / chloramphenicol vector (pSB1AC3) and I13600 is on an Ampicillin vector, we decided to first transform I13600 into a cell and select for Ampicillin. We then checked for CFP expression and made the cells competent. After this we transformed K182005 into these cells and selected for Chloramphenicol.<br> <br> | Since K182005 is on an Ampicillin / chloramphenicol vector (pSB1AC3) and I13600 is on an Ampicillin vector, we decided to first transform I13600 into a cell and select for Ampicillin. We then checked for CFP expression and made the cells competent. After this we transformed K182005 into these cells and selected for Chloramphenicol.<br> <br> | ||
| − | With both plasmids now in the cell we checked for fluorescence as can be seen in the picture below. <br><br> | + | With both plasmids now in the cell we checked for fluorescence, in the presence and absence of anhydrotetracycline (AHT or ATc), as can be seen in the picture below. <br><br> |
[[Image:4 K182005-I13600 flu no AHTAberdeen2009.jpg|center|400 px]] | [[Image:4 K182005-I13600 flu no AHTAberdeen2009.jpg|center|400 px]] | ||
Revision as of 22:17, 21 October 2009
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_I13600
- Grew up this Biobrick on a plate just to test its flourescence compared to the other constitutive flourescent proteins in the Registry as of July 2006. This has one of the weakest flourescent outputs. --Smelissali 19:39, 3 July 2006 (EDT)
User Reviews
UNIQc39c5e280265afbc-partinfo-00000000-QINU
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Smelissali |
Brick works, but the cyan flourescent protein is definitely not the brightest reporter you could use, and thus not my first choice. --Smelissali 19:38, 3 July 2006 (EDT) |
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Antiquity |
This review comes from the old result system and indicates that this part worked in some test. |
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Aberdeen_Scotland 2009 |
The gel analysis and the plasmid miniprep worked quite nicely as expected. But later the construct was further tested and characterized by combining with the bio-brick BBa_K182005 which contained a tet repressor under the control of a CI operator. So, the CFP expression of BBa_I13600was repressed as expected when tested with the BBa_K182005. With the addition of Anhydrotetracycline (an analogue of tetracycline) the CFP expression was restored and this was confirmed performing the fluorescent microscopy assay.
 Transforming BBa_K182005 into these cells should stop the production of CFP, since it expresses TetR. If it works as anticipated, the K182005 Tet repressor should bind to the Tet Operator of I13600 which would stop the expression of CFP.
With both plasmids now in the cell we checked for fluorescence, in the presence and absence of anhydrotetracycline (AHT or ATc), as can be seen in the picture below. 
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UNIQc39c5e280265afbc-partinfo-00000005-QINU