Team:Amsterdam/data/experimental

From 2012.igem.org

(Difference between revisions)
Line 28: Line 28:
[[File:Amsterdam_exp_fig_4.png|300px|right|thumb|Figure 3]]
[[File:Amsterdam_exp_fig_4.png|300px|right|thumb|Figure 3]]
We aimed to characterize the activity of our writer during bacterial growth.  We performed a growth curve experiment (figure 3) and collected samples at different time points to test the occurrence of methylation over different times of growth.  
We aimed to characterize the activity of our writer during bacterial growth.  We performed a growth curve experiment (figure 3) and collected samples at different time points to test the occurrence of methylation over different times of growth.  
-
[[File:Amsterdam_exp_fig_3.png|300px|right|thumb|Figure 4]]
+
[[File:Amsterdam_exp_fig_5.png|300px|right|thumb|Figure 4]]
Figure 4 shows ScaI digestion of plasmids extracted from samples activated or not by 10mM IPTG and collected between 0 and 12h. Both conditions reveal an intermediate restriction profile over. However, the two samples taken after 24 hours show a different result, i.e. almost only uncut plasmid is observed as a result of an significant increase of methylation of our reader.   
Figure 4 shows ScaI digestion of plasmids extracted from samples activated or not by 10mM IPTG and collected between 0 and 12h. Both conditions reveal an intermediate restriction profile over. However, the two samples taken after 24 hours show a different result, i.e. almost only uncut plasmid is observed as a result of an significant increase of methylation of our reader.   
Line 35: Line 35:
The same bacterial batch was used in both experiments but for the stationary experiment first grown over a longer period of time (at least more than 24 hours), either with or without the inducer.  
The same bacterial batch was used in both experiments but for the stationary experiment first grown over a longer period of time (at least more than 24 hours), either with or without the inducer.  
-
 
+
[[File:Amsterdam_exp_fig_3.png|300px|right|thumb|Figure 5]]
-
[[File:Amsterdam_exp_fig_5.png|300px|right|thumb|Figure 5]]
+
Figure 5 shows the restriction profile taken from three different time points: 48, 72 and 96 hours. The first three restriction profiles correspond to growth without IPTG induction and the last three with 10 mM IPTG. During the culture no additional IPTG was added since we expect that IPTG is not degraded over time. We do not observe a change in the restriction profile, meaning that the time of culture does not influence our results.  
Figure 5 shows the restriction profile taken from three different time points: 48, 72 and 96 hours. The first three restriction profiles correspond to growth without IPTG induction and the last three with 10 mM IPTG. During the culture no additional IPTG was added since we expect that IPTG is not degraded over time. We do not observe a change in the restriction profile, meaning that the time of culture does not influence our results.  

Revision as of 03:25, 27 September 2012