Difference between revisions of "Part:BBa K2145000:Experience"
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[[File:Splates95102.png]]<br> | [[File:Splates95102.png]]<br> | ||
| − | Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see | + | Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG95 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR. |
[[File:Alverno_ca_a95.png]] | [[File:Alverno_ca_a95.png]] | ||
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[[File:Alverno_ca_g_95.png]] | [[File:Alverno_ca_g_95.png]] | ||
| − | Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones | + | Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones containing this part in liquid culture. Again, we emphasize that only RFP OR GFP are expressed, but never both. We believe this may be due to supercoiling -- when one gene expresses, it creates supercoils that shut down the other gene. |
Sequencing on this plasmid was low-quality and inconclusive. | Sequencing on this plasmid was low-quality and inconclusive. | ||
Latest revision as of 00:31, 31 October 2016
Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG95 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.
Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones containing this part in liquid culture. Again, we emphasize that only RFP OR GFP are expressed, but never both. We believe this may be due to supercoiling -- when one gene expresses, it creates supercoils that shut down the other gene.
Sequencing on this plasmid was low-quality and inconclusive.
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