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Minnesota-experimental/8 July 2009
From 2009.igem.org
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- Samples were collected from DH5αPro cells containing the TNN and TTN promoter constructs mutant at position 4. Aliquots of culture were collected and fixed 3, 6, 9, and 12 hours following innoculation of the cultures in media containing various concentrations of aTc and IPTG as inducers. TOP10 cells containing the same constructs were also sampled as a control.
- Transformed TOP10 cells containing promoter constructs mutant at position 6 were screened using a fluorescent camera to select colonies producing GFP. This indicated that the cells within that colony contained plasmids with active promoters.
- Colonies which appeared green were transferred to a second antibiotic LB agar plate for further screening. The plate was marked into a numbered grid, and individual colonies were transferred into each square using a sterile micropipette tip. Cells were allowed to grow at 37 °C overnight.
