Difference between revisions of "Team:Birkbeck/Results"

 
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<IMG SRC="https://static.igem.org/mediawiki/2015/e/e8/Birkbeck_150916_LP_gel1_pred.png" height="600px" width="300px">
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<p>a.&nbsp;<IMG SRC="https://static.igem.org/mediawiki/2015/e/e8/Birkbeck_150916_LP_gel1_pred.png" height="600px" width="300px">
<IMG SRC="https://static.igem.org/mediawiki/2015/1/18/Birkbeck_150916_LP_gel1.jpeg" height="600px" width="350px">
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b.&nbsp;<IMG SRC="https://static.igem.org/mediawiki/2015/1/18/Birkbeck_150916_LP_gel1.jpeg" height="600px" width="350px"></p>
<p>Comparison of predicted and actual band sizes on gel 1.</p>
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<p><b>Fig 1.</b> Comparison of predicted (a) and actual (b) band sizes on gel 1.</p>
 
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<IMG SRC="https://static.igem.org/mediawiki/2015/6/60/Birkbeck_150916_LP_gel2_pred.png" height="600px" width="300px">
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<p>a.&nbsp;<IMG SRC="https://static.igem.org/mediawiki/2015/6/60/Birkbeck_150916_LP_gel2_pred.png" height="600px" width="300px">
<IMG SRC="https://static.igem.org/mediawiki/2015/f/f9/Birkbeck_150916_LP_gel2.jpeg" height="600px" width="350px">
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b.&nbsp;<IMG SRC="https://static.igem.org/mediawiki/2015/f/f9/Birkbeck_150916_LP_gel2.jpeg" height="600px" width="350px"></p>
<p>Comparison of predicted and actual band sizes on gel 2.
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<p><b>Fig 2.</b> Comparison of predicted (a) and actual (b) band sizes on gel 2.</p>
</p>
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<h2><b>Results</b></h2>
 
 
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            <h3>
 
                <a href="#">InterLab Study Results
 
            </h3>
 
                    <p>Click here to see the results of our contribution to iGEM's InterLab Study, and some additional experiments that support our research.
 
                    </p>   
 
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<p><IMG SRC="https://static.igem.org/mediawiki/2015/5/58/Growth_Curve_50_mL_600_nm_team_birkbeck_iGEM_2015.jpg"></center>
 
<p><b><u>Fig. 1: Growth Curve of <i>E. coli</i> DH5α Strains Following Culture Optical Density of 600 nm</u></b>.</p>
 
 
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<p>Growth kinetics was initially studied using 50 mL cultures. <b>Fig. 1</b> shows the growth kinetics of <i>E. coli</i> DH5α & derivative strains containing plasmids from the <a href="https://2015.igem.org/Team:Birkbeck/InterLab_Study">InterLab</a> study.  The growth curve shows that the <i>E. coli</i> strain that contains the P1-<i>gfp</i> expression device grows at a slower rate than the other strains investigated. At 220 minutes the <i>E. coli</i> DH5α P1 strain has a significantly lower OD<sub>600</sub> than the <i>E. coli</i> DH5α (P=0.023). <i>E. coli</i> DH5α remains significantly higher in OD<sub>600</sub> than <i>E. coli</i> DH5α with the P1-<i>gfp</i> expression device (P=<0.001)<!---->. The only difference between the <i>E. coli</i> DH5α & <i>E. coli</i> DH5α positive control device is observed at 280 minutes into the growth curve (P=0.016) where the positive control has a higher OD<sub>600</sub>. The multiple comparison table showing P values can be viewed in <a href="https://2015.igem.org/Team:Birkbeck/Results/Table_S1"><b>Table S1</b>.</a></p>.
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<!--button for Conclusion-->
<center><img src="https://static.igem.org/mediawiki/2015/a/a8/Growth_Curve_%28395_nm%29_50_mL_Cultures_Team_birkbeck_iGEM_2015.jpg "></center>
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<a href="https://2015.igem.org/Team:Birkbeck/Conclusion"><img width="400px" height="auto" src="https://static.igem.org/mediawiki/2015/7/7d/Birkbeck_phage_infection2.png" border="0"/></a>
<p><b><u>Fig. 2: Growth Curve of <i>E. coli</i> DH5α Strains Following Culture Optical Density of 395 nm</u></b>.</p>
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<p>To find concluding remarks about our project</p>
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<!--button for Research-->
<p>In order to investigate if there could be a point in the <i>E. coli</i> DH5α growth curve in which a signal from the GFP could be detected by absorbance, the growth curves were also conducted using the major absorption peak of GFP (wavelength 395 nm)<!--(REF!)-->. The growth curve data for the culture optical density is displayed in <b>Fig. 2</b>.</P>
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<a href="https://2015.igem.org/Team:Birkbeck/Research"><img width="400px" height="auto" src="https://static.igem.org/mediawiki/2015/c/c7/Rachel-elliot.jpg" border="0"/></a>
<p>When comparing the data point of <i>E. coli</i> DH5α strains, there appears to be a significant increase in culture OD<sub>395</sub> in the <i>E. coli</i> cells with the P1-<i>gfp</i> expression device (P<0.001). This apparent signal is only present between 60-100 minutes of growth. When comparing the positive control & <i>E. coli</i> DH5α1, there is no significant difference between the data sets at 60 or 100 minutes (P=1 for both time points). A small potential signal is observed from the oositive control <i>gfp</i> expression device at 280 minutes (P=0.012) & 300 minutes (P=0.006). This significance is lost after 300 minutes (P=0.262). See <a href="https://2015.igem.org/Team:Birkbeck/Results/Table_S2"><b>Table S2</b></a> for more details. In order to verify these results & to test for the feasibility of scaling down to a 96-well microtitre plate assay, a 10 hour growth curve was conducted in a 96-well microtitre plate (see <b>Fig. 6-8</b>).</p>
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<p>Back to main Research page</p>
<!--Multiple Comparison table link for OD 395 nm (table S2); https://2015.igem.org/Team:Birkbeck/Results/Table_S2. issue sorted, this page is good to go!.-->
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<center><img src="https://static.igem.org/mediawiki/2015/d/d7/Viable_count_1_hour_%28dh5_alpha%29_team_birkbeck_iGEM_2015.jpg"></center>
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<p><b><u>Fig. 3: Viable Count of <i>E. coli</i> DH5α After 60 mins</u></b>.</p>
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<p>In order to assess how many viable cells correspond to different optical densities, a viable count was conducted on <i>E. coli</i> DH5α1 at 60 minutes (<b>Fig. 3</b> & <b>Table 1</b>), 175 minutes (<b>Fig. 5</b> & <b>Table 2</b>) & 225 minutes (data not shown due to high level of contamination).</p>
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<p>Considering the OD<sub>600</sub> of the <i>E. coli</i> DH5α1 cultures at 60 mins, triplicate cultures were OD<sub>600</sub> = 0.029, 0.01 & 0.025<!--0.021 mean-->. The viable count of each of the cultures gave a mean of 2.57×10<sup>6</sup> cfu/mL (<b>Table 1</b>). It can therefore be concluded that an <i>E. coli</i> DH5α culture at an OD<sub>600</sub> = 0.021 corresponds to approximately 2.57×10<sup>6</sup> cfu/mL.</p>
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<caption class="title" style="width:522px">Descriptive Statistics of 1 hour Viable Count of <i>E. coli</i> DH5α.<span class="details"></span></caption>
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<td class="cornerLabels">&nbsp;</td>
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<td class="columnLabels dataAreaLeft vCC role3">N</td>
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<td class="columnLabels vCC role3">Std. Deviation</td>
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<td class="rowLabels dataAreaTop role3">Viable Count</td>
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<td class=" e dataAreaTop dataAreaLeft vCC">9</td>
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<td class=" e dataAreaTop vCC">1600000</td>
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<td class=" e dataAreaTop vCC">3750000</td>
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<td class=" e dataAreaTop vCC">2566666.67</td>
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<td class=" e dataAreaTop vCC">627495.020</td>
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<td class="rowLabels hCR role3">Valid N (listwise)</td>
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<td class=" o dataAreaLeft hCR vCC">9</td>
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<td class=" o hCR vCC">&nbsp;</td>
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<!--End of descriptive Stats table-->
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<p><b><u>Table 1: Descriptive Statistics of 60 minutes Viable Count of <i>E. coli</i> DH5α.</u></b>.</p>
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<br>
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<center><img src="https://static.igem.org/mediawiki/2015/e/e6/Viable_count_of_DH5_alpha_after_175_mins_team_birkbeck_iGEM_2015.jpg"></center>
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<p><b><u>Fig. 4: Viable Count of <i>E. coli</i> DH5α After 175 mins</u></b>.</p>
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<br>
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<p>At 175 minutes into growth, the <i>E. coli</i> DH5α1 cultures had OD<sub>600</sub> of; 0.255, 0.216 & 0.262<!--mean = 0.244-->. The viable count of each of the cultures gave a mean of 1.33×10<sup>8</sup> cfu/mL (<b>Table 2</b>). It can therefore be concluded that an <i>E. coli</i> DH5α culture at an OD<sub>600</sub> = 0.244 corresponds to approximately 1.33×10<sup>8</sup> cfu/mL. Considering the previous OD<sub>600</sub> (0.021), there is approximately a 10-fold increase in OD<sub>600</sub> which corresponds to nearly a 100-fold increase in viable cells.</p>
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<!--Begining of the descriptive statistics table for 175 minutes of growth--> </p>
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<caption class="title" style="width:547px"><span class="details">Descriptive Statistics of 175 Minutes Viable Count of <i>E. coli</i> DH5α.</span></caption>
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<td class="cornerLabels">&nbsp;</td>
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<td class="columnLabels dataAreaLeft vCC role3">N</td>
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<td class="columnLabels vCC role3">Minimum</td>
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<td class="columnLabels vCC role3">Maximum</td>
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<td class="columnLabels vCC role3">Mean</td>
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<td class="columnLabels vCC role3">Std. Deviation</td>
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<td class="rowLabels dataAreaTop role3">Viable Count</td>
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<td class=" e dataAreaTop dataAreaLeft vCC">9</td>
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<td class=" e dataAreaTop vCC">90000000</td>
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<td class=" e dataAreaTop vCC">175000000</td>
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<td class=" e dataAreaTop vCC">133333333.33</td>
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<td class=" e dataAreaTop vCC">29154759.474</td>
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<td class="rowLabels hCR role3">Valid N (listwise)</td>
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<td class=" o dataAreaLeft hCR vCC">9</td>
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<td  style="width:114px"></td>
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<td  style="width:67px"></td>
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<td  style="width:83px"></td>
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<td  style="width:91px"></td>
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<td  style="width:96px"></td>
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<td  style="width:96px"></td>
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<!--End of the descriptive statistics table for 175 minutes of growth-->
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<p><b><u>Table 2: Descriptive Statistics of 175 minutes Viable Count of <i>E. coli</i> DH5α.</u></b></p>
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<br>
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<center><IMG SRC="https://static.igem.org/mediawiki/2015/8/8a/Growth_curve_601_nm_96-microtitre_Team_birkbeck_iGEM_2015_data.jpg "></center>
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<p><b><u>Fig. 5: Growth Curves of Different Strains of <i>E. coli</i> DH5α Following Culture Optical Density at 601 nm</u></b>.</p>
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<br>
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<p>All data analysis tables for the OD<sub>601</sub> growth curves are in; <a href="https://2015.igem.org/Team:Birkbeck/table_S3">Table S3</a>, <a href="https://2015.igem.org/Team:Birkbeck/table_S5">Table S5</a> & <a href="https://2015.igem.org/Team:Birkbeck/table_S6">Table S6</a> for 0-200 minutes, 220-420 minutes & 440-580 minutes respectively. Comparing the growth of <i>E. coli</i> DH5α containing P1-<i>gfp</i> expression device with the <i>E. coli</i> DH5α, there is no significant difference between the two cultures at 80 minutes (P=0.943), however, at 100 minutes there is a very highly significant difference between the 2 strains of <i>E. coli</i> DH5α (P<0.001). The OD<sub>601</sub> of <i>E. coli</i> DH5α containg the P1-<i>gfp</i> expression device remains at a significantly lower that the <i>E. coli</i> DH5α untill 400 minutes (P=0.441).</p>
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<p>Comparing the <i>E. coli</i> DH5α P1-<i>gfp</i> expression device with the positive control <i>gfp</i> <i>E. coli</i> DH5α cultures, the OD<sub>601</sub> remains insignificantly different at 100 minutes (P=0.848) with significance being observed at 120 minutes (P<0.001). The significance is observed until 280 minutes into the growth curves (P=0.065). Interesting;ly, this is a shorter window of significance compared to <i>E. coli</i> P1-<i>gfp</i> expression device & <i>E. coli</i> DH5α (160 minute Vs 300 minutes respectively).</p>
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<p>Comparing P1 & P2-<i>gfp</i> expression devices in <i>E. coli</i> DH5α, no significance is observed at 100 minutes (P=0.132). P1-<i>gfp</i> expression device is very highly significantly lower (P<0.001) than P2-<i>gfp</i> expression device after 120 minutes of culturing. This significance is observed until 320 minutes of culturing (P=0.075).</p>
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<p>When comparing the positive control, P2-<i>gfp</i> & <i>E. coli</i> DH5α, there is no significant difference throughout the growth curve. After 420 minutes, there is no significant difference between any of the <i>E. coli</i> DH5α cultures OD<sub>601</sub> (P=0.81).</p>
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<!--Table S3 (0-200 mins); https://2015.igem.org/Team:Birkbeck/table_S3, (change to S4!!!)Tables S5 (220-420); https://2015.igem.org/Team:Birkbeck/table_S5, Table S6 (440-580 minutes); https://2015.igem.org/Team:Birkbeck/table_S6  -->
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<br>
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<center><IMG SRC="https://static.igem.org/mediawiki/2015/d/d0/Growth_curve_501_nm_microtitre_data_Team_birkbeck_iGEM_2015.jpg"></center>
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<p><b><u>Fig. 6: Growth Curves of Different Strains of <i>E. coli</i> DH5α Following Culture Optical Density at 501 nm</u></b>.</p>
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<br>
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<p>All data analysis tables for the OD<sub>501</sub> growth curves are in; <a href="https://2015.igem.org/Team:Birkbeck/table_S7">Table S7</a>, <a href="https://2015.igem.org/Team:Birkbeck/table_S8">Table S8</a> & <a href="https://2015.igem.org/Team:Birkbeck/table_S9">Table S9</a> for 0-200 minutes, 220-420 & 440-580 minutes respectively. Comparing the growth of <i>E. coli</i> DH5α containing P1-<i>gfp</i> expression device with the <i>E. coli</i> DH5α, there is no significant difference between the two cultures at 80 minutes (P=0.987). After 100 minutes there is a highly significant difference between the two cultures (P=0.003). This difference remains significant until 400 minutes into culturing (P=0.093).</p>
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<p>Comparing the <i>E. coli</i> DH5α P1-<i>gfp</i> expression device with the positive control <i>gfp</i> <i>E. coli</i> DH5α cultures, the OD<sub>501</sub> remains insignificantly different at 100 minutes (P=0.85) with significance being observed at 120 minutes (P<0.001). The P1-<i>gfp</i> expression device culture remains significantly lower than the positive control <i>gfp</i> expression device until 320 minutes (P=0.053).</p>
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<p>Comparing P1 & P2-<i>gfp</i> expression devices in <i>E. coli</i> DH5α, no significance is observed at 100 minutes (P=0.403). P1-<i>gfp</i> expression device is very highly significantly lower (P<0.001) than P2-<i>gfp</i> expression device after 120 minutes of culturing. This significance is observed until 320 minutes of culturing (P=0.096).</p>
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<p>When comparing the positive control, P2-<i>gfp</i> & <i>E. coli</i> DH5α, there is no significant difference throughout the growth curve. After 420 minutes, there is no significant difference between any of the <i>E. coli</i> DH5α cultures OD<sub>501</sub> (P=0.09). These results are identical to the results obtained in th OD<sub>501</sub> absorption of the <i>E. coli</i> DH5α cultures.</p>
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<!--Table S7 (0 mins - 200 mins); https://2015.igem.org/Team:Birkbeck/table_S7 , Table S8 (240-420 minutes); https://2015.igem.org/Team:Birkbeck/table_S8 Table S9 (440-580 mins); https://2015.igem.org/Team:Birkbeck/table_S9-->
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<br>
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<center><img src="https://static.igem.org/mediawiki/2015/4/4a/Growth_curve_475_nm_microtitre_team_birkbeck_iGEM_2015.jpg"></center>
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<p><b><u>Fig. 7: Growth Curves of Different Strains of <i>E. coli</i> DH5α Following Culture Optical Density at 475 nm</u></b>.</p>
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<br>
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<p>All data analysis tables for the OD<sub>475</sub> growth curves are in; <a href="https://2015.igem.org/Team:Birkbeck/Table_S10">Table S10</a>, <a href="https://2015.igem.org/Team:Birkbeck/table_S11">Table S11</a> & <a href="https://2015.igem.org/Team:Birkbeck/table_S12">Table S12</a> for 0-200 minutes, 220-420 & 440-580 minutes respectively. Comparing the growth of <i>E. coli</i> DH5α containing P1-<i>gfp</i> expression device with the <i>E. coli</i> DH5α, there is no significant difference between the two cultures at 100 minutes (P=0.982). After 120 minutes there is a highly significant difference between the two cultures (P=0.003). This difference remains significant until 440 minutes into culturing (P=0.745).</p>
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<p>Comparing the <i>E. coli</i> DH5α P1-<i>gfp</i> expression device with the positive control <i>gfp</i> <i>E. coli</i> DH5α cultures, the OD<sub>475</sub> remains insignificantly different at 100 minutes (P=0.844) with significance being observed at 120 minutes (P<0.001). The P1-<i>gfp</i> expression device culture remains significantly lower than the positive control <i>gfp</i> expression device until 340 minutes (P=0.19).</p>
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<p>Comparing P1 & P2-<i>gfp</i> expression devices in <i>E. coli</i> DH5α, no significance is observed at 100 minutes (P=0.464). P1-<i>gfp</i> expression device is very highly significantly lower (P<0.001) than P2-<i>gfp</i> expression device after 120 minutes of culturing. This significance is observed until 340 minutes of culturing (P=0.163).</p>
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+
<p>When comparing the positive control, P2-<i>gfp</i> & <i>E. coli</i> DH5α, there is no significant difference throughout the growth curve. Conducting a one-way ANOVA of the culture density at 440+ minutes shows there is a highly significant difference between the means (P=0.003). The multiple comparisons table shows no significant difference between any of the individual data point (data not shown).</p>
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+
<!--Table S10 (0 mins-200 mins); https://2015.igem.org/Team:Birkbeck/Table_S10 Table S11 (220-420); https://2015.igem.org/Team:Birkbeck/table_S11 Table S12 (440-580 minutes); https://2015.igem.org/Team:Birkbeck/table_S12, P1 remains statistically lower than DH5 until 440 mins (P=0.745) giving the P=0 value.-->
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<br>
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<center><img src="https://static.igem.org/mediawiki/2015/8/8b/Growth_curve_395_microtitre_team_birkbeck_iGEM_2015.jpg "></center>
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<p><b><u>Fig. 8: Growth Curves of Different Strains of <i>E. coli</i> DH5α Following Culture Optical Density at 395 nm</u></b>.</p>
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<br>
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<p>All data analysis tables for the OD<sub>395</sub> growth curves are in; <a href="https://2015.igem.org/Team:Birkbeck/Table_S13">Table S13</a>, <a href="https://2015.igem.org/Team:Birkbeck/table_S14">Table S14</a> & <a href="https://2015.igem.org/Team:Birkbeck/table_S15">Table S15</a> for 0-200 minutes, 220-420 & 440-580 minutes respectively. Comparing the growth of <i>E. coli</i> DH5α containing P1-<i>gfp</i> expression device with the <i>E. coli</i> DH5α, there is no significant difference between the two cultures at 80 minutes (P=0.937). After 100 minutes there is a highly significant difference between the two cultures (P<0.001). This difference remains significant until 440 minutes into culturing (P=0.083)</p>
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<p>Comparing the <i>E. coli</i> DH5α P1-<i>gfp</i> expression device with the positive control <i>gfp</i> <i>E. coli</i> DH5α cultures, the OD<sub>395</sub> remains insignificantly different at 80 minutes (P=0.992) with significance being observed at 100 minutes (P=0.011). The P1-<i>gfp</i> expression device culture remains significantly lower than the positive control <i>gfp</i> expression device until 440 minutes (P=0.325).</p>
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<p>Comparing P1 & P2-<i>gfp</i> expression devices in <i>E. coli</i> DH5α, no significance is observed at 100 minutes (P=0.358). P1-<i>gfp</i> expression device is very highly significantly lower (P<0.001) than P2-<i>gfp</i> expression device after 120 minutes of culturing. This significance is observed until 340 minutes of culturing (P=0.171).</p>
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<p>When comparing the positive control, P2-<i>gfp</i> & <i>E. coli</i> DH5α, there is no significant difference throughout the growth curve. Conducting a one-way ANOVA of the culture density at 440+ minutes shows there is a highly significant difference between the means (P<0.001). The multiple comparisons table shows no significant difference between any of the individual data point (data not shown).</p>
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<!--Table S13 (0-200 minutes); https://2015.igem.org/Team:Birkbeck/Table_S13 Table S14 (220-420 mins); https://2015.igem.org/Team:Birkbeck/table_S14 Table S15 (440-580 mins); https://2015.igem.org/Team:Birkbeck/table_S15 (the P1 is not significant at 440 minutes [P=0.083]-->
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<br>
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<center><img src="https://static.igem.org/mediawiki/2015/9/9a/Fluorescence_growth_curve_of_multiple_strains_of_E._coli_DH5_alpha_team_Birkbeck_iGEM_2015.jpg "></center>
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<p><b><u>Fig. 9: Growth Curves of Different Strains of <i>E. coli</i> DH5α Following Culture Fluorescence</u></b>.</p>
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<br>
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<p>All data analysis tables for the culture fluorescence growth curves are; <a href="https://2015.igem.org/Team:Birkbeck/table_S16">Table S16</a>, <a href="https://2015.igem.org/Team:Birkbeck/Table_S17 ">Table S17</a> & <a href="https://2015.igem.org/Team:Birkbeck/table_S18">Table S18</a> for 0-200 minutes, 220-420 & 440-580 minutes respectively. Comparing the P1-<i>gfp</i> expression device with <i>E. coli</i> DH5α, at 0 minutes there is a highly significant difference between the two cultures fluorescence (P=0.005). This significance is observed until 100 minutes (P=0.427). There is an insignificant difference between the two cultures until 160 minutes (P<0.001). At 220 minutes into the growth curve, the fluorescence between P1<i>gfp</i> expression device is insignificant (P=0.277) with significance being observed at 240 minutes (P=0.001). The fluorescence of P1-<i>gfp</i> expression device remains higher than the <i>E. coli</i> DH5α culture.</p>
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<p>Comparing the positive control <i>gfp</i> expression device with <i>E. coli</i> DH5α, at no point in the growth curve does the fluorescence of the positive control <i>gfp</i> expression device increase above that of the <i>E. coli</i> DH5α culture. Even at 580 minutes of culturing the positive control does not increase in fluorescence above the <i>E. coli</i> <i>E. coli</i> DH5α (P=0.677).</p>
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<p>Comparing the P2-<i>gfp</i> expression device with <i>E. coli</i> DH5α, Initially there is no significant difference between the two cultures in fluorescence (P=1). There is a highly significant Increase in fluorescence from P2 at 180 minutes (P=0.007). At 200 minutes, there is no significant difference in fluorescence observed between P2 + <i>E. coli</i> DH5α (P=0.146). Insignificance of fluorescence between the two cultures is observed until 500 minutes of culturing (P=0.038). The fluorescence from the P2-<i>gfp</i> expression device remains significantly higher than <i>E. coli</i> DH5α throughout the rest of the growth curve.</p>
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<p>Comparing the P1-<i>gfp</i> expression device with the positive control <i>gfp</i> expression device there is a significantly higher fluorescence from 0 minutes (P=0.034) & 20 minutes (P=0.038). At 40 minutes of culturing there is no significant difference in fluorescence (P=0.164). At 60 minutes the P1-<i>gfp</i> expression device has a significantly higher fluorescence than the positive control (P=0.017). The significance is not observed at 80 minutes (P=0.849). There is no significant fluorescence from P1 compared to the positive control until 180 minutes (P=0.009). No significance is observed between the two promoters until 340 minutes where P1 has a higher fluorescence (P=0.019). The P1-<i>gfp</i> expression device has a higher fluorescence compared to the positive control at 360 minutes (P=0.024) and becomes insignificant at 380 minutes (P=0.245) where it remains insignificant for the result of the culturing time.</p>
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<p>Comparing P2-<i>gfp</i> expression device with the positive control, there is no time point where the P2-<i>gfp</i> expression device has a statistically higher fluorescence than the positive control <i>gfp</i> expression device.</p>
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<p>When comparing the data sets for the P1 & P2 <i>gfp</i> expression devices, there is no significant difference until 60 minutes (P=0.021). At 60 minutes, the P1-<i>gfp</i> expression device has a higher fluorescence. There is no significance between P1 & P2 <i>gfp</i> expression devices at 80 minutes (P=0.646) & no other time points show significance between these two cultures.</p>
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<br><hr><br>
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<p>Results discussion & credit for the work are shown in the <a href="https://2015.igem.org/Team:Birkbeck/Discussion">Discussion</a> section of this website.</p>
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<!--For the fluorescence data, the table has to be split into 3 different pages! bit of a ball-ache but it has to be done its just a really big file. I am guessing the same has to be done with the absorption spectra data :( lots more work needs doing on this matter. Table S16 (0-200); https://2015.igem.org/Team:Birkbeck/table_S16 Table S17 (220-420); https://2015.igem.org/Team:Birkbeck/Table_S17 Table S18 (440 - 580 minutes); https://2015.igem.org/Team:Birkbeck/table_S18-->
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<a href="https://2015.igem.org/Team:Birkbeck/Conclusion">Up next: Conclusions</a>
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Latest revision as of 22:42, 18 September 2015

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Results

BioBrick Cloning Results

Click here to see the results from testing the constructs ORF-314, stf gene, tfa construct, P(Cat)-TetR construct, TetR-controlled tfa circuit and cI-Cro circuit.


To confirm the presence and correct size of the BioBricks we are submitting, all samples were run on an agarose gel. The constructs tested were:

  • ORF-314
  • the stf gene
  • the tfa construct
  • the P(Cat)-TetR construct
  • the TetR-controlled tfa circuit
  • the cI-Cro circuit

All samples were initially digested with two of the four standard iGEM restriction enzymes (XbaI, EcoRI, SpeI and PstI) and expected fragment sizes calculated (Fig 1a and 2a). Success of the cloning was then determined by comparing actual band sizes to the predicted values (Fig 1b and 2b).


a.  b. 

Fig 1. Comparison of predicted (a) and actual (b) band sizes on gel 1.


a.  b. 

Fig 2. Comparison of predicted (a) and actual (b) band sizes on gel 2.





To find concluding remarks about our project

Back to main Research page