Difference between revisions of "Part:BBa K2145103:Experience"
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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 GG108 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. | 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 GG108 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. | ||
Revision as of 23:50, 30 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 GG108 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.
Above is TX-TL fluorescence data for this set of plasmids. Positive control is a GFP gene on a strong promoter and strong RBS (pBEST-OR2-OR1-Pr-UTR1-deGFP-T500, available on addgene). This plasmid did NOT express either RFP or GFP in TX-TL when added at 2nM (with dCas expression plasmid and gRNA expression plasmid). Addition of inducers to TX-TL reactions with these plasmids did not affect expression.
Sequencing on this plasmid was low-quality and inconclusive.
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