Team:Amsterdam/data/experimental

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The third characterisation experiment concerns the stimulation of pSB1AT3/pBad/Mtase in Efficient® DH5α™ competent cells (Invitrogen) in stationary phase with addition of 2% arabinose daily. Samples were taken after 48, 72 and 96 hours, and digested with ScaI restriction enzyme (figure 9). These results are consistent with the previous results, showing a gradual shift towards the “on-state” with induction of the pBad promoter by addition of arabinose daily.
The third characterisation experiment concerns the stimulation of pSB1AT3/pBad/Mtase in Efficient® DH5α™ competent cells (Invitrogen) in stationary phase with addition of 2% arabinose daily. Samples were taken after 48, 72 and 96 hours, and digested with ScaI restriction enzyme (figure 9). These results are consistent with the previous results, showing a gradual shift towards the “on-state” with induction of the pBad promoter by addition of arabinose daily.
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[[File:Amsterdam_exp_fig_7.png|300px|right|thumb|Figure 10]]
The “off” state was not achieved in the absence of arabinose. All the experiments have been conducted into the high copy number plasmid pSB1AT3. It is known that leaky expression is relative to the copy number of the plasmid.1 Hence, the inability of the Cellular Logbook to show an “off” state in the absence of arabinose could be attributed to the high copy number plasmid used in these experiments as was discussed with the LacH.
The “off” state was not achieved in the absence of arabinose. All the experiments have been conducted into the high copy number plasmid pSB1AT3. It is known that leaky expression is relative to the copy number of the plasmid.1 Hence, the inability of the Cellular Logbook to show an “off” state in the absence of arabinose could be attributed to the high copy number plasmid used in these experiments as was discussed with the LacH.
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Similarly to the pSB1AT3/LacH/Mtase, a growth curve experiment was conducted to characterize the acquired construct of pSB1AT3/pBad/Mtase.
Similarly to the pSB1AT3/LacH/Mtase, a growth curve experiment was conducted to characterize the acquired construct of pSB1AT3/pBad/Mtase.
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[[File:Amsterdam_exp_fig_7.png|300px|right|thumb|Figure 10]]
 
Over the course of time the methylation-dependent restriction profile observed in the gel showed a shift towards the ‘on’ digestion profile in the presence of arabinose. This result shows that our Cellular Logbook is able to sense and write an arabinose signal present in the medium in a shorter period of time.
Over the course of time the methylation-dependent restriction profile observed in the gel showed a shift towards the ‘on’ digestion profile in the presence of arabinose. This result shows that our Cellular Logbook is able to sense and write an arabinose signal present in the medium in a shorter period of time.

Revision as of 02:42, 27 September 2012