Team:Austin Texas/photocage
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Once these components were assembled by Gibson Assembly, the two plasmids were transformed via electroporation into aliquots of Amberless E.coli. | Once these components were assembled by Gibson Assembly, the two plasmids were transformed via electroporation into aliquots of Amberless E.coli. | ||
- | After transformation, the amberless E. coli cells containing both the synthetase/tRNA pair plasmid and the amber T7 RNAP/GFP plasmid were assayed for their ability to express GFP upon irradiation | + | After transformation, the amberless E. coli cells containing both the synthetase/tRNA pair plasmid and the amber T7 RNAP/GFP plasmid were assayed for their ability to express GFP upon irradiation with 365 nm light in different intervals of time. The cells were irradiated for 0 minutes, 1 minute, 5 minutes, 15 minutes, and 30 minutes in order to observe the temporal control of GFP expression. |
After irradiation with light, these cells were allowed to grow overnight before taking fluorescent measurements. This additional growth was necessary to allow the "decaged" T7 RNA polymerase to polymerase mRNA transcripts of the GFP coding sequence. | After irradiation with light, these cells were allowed to grow overnight before taking fluorescent measurements. This additional growth was necessary to allow the "decaged" T7 RNA polymerase to polymerase mRNA transcripts of the GFP coding sequence. |
Revision as of 23:20, 14 October 2014
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