Team:Alverno CA/Results

Alverno iGEM 2016

Results













GG28_1-5

This set of plasmids is assembled so that the RFP and GFP inducers are convergent (Pointing towards each other). The time course fluorescence traces from 1000-bp spacer constructs from our plate reader measured the fluorescence of RFP and GFP in the bacteria. In figure 1:

Figure 1

This graph shows the fluorescence of RFP within the plasmids. (D01, E01,F01, G01, and H01 are all clones of the same plasmid). As seen in the graph, there is a strange range of growth in bacteria, which was present in almost all of our results from TX-TL.

Figure 2

Figure 2 shows the plate reader’s measurement of the fluorescence of GFP in our bacteria. (D12, E12, F12, G12, H12 are all clones of the same plasmid)  This graph shows results that are similar to that of RFP, however, the overall fluorescence levels are higher than the fluorescence levels in RFP.
 

Figure 3

These strains have been transformed with both RFP and GFP (A1, A2, A3, A4, A5 are all clones of the same plasmid with both inducers. ) The fluorescence traces shown in this graph shows that overall the bacteria was successfully transformed, however, this data is not consistent with the data from the two previous graphs (those specific to RFP and GFP respectively).

Figure 4


Figure 5

Figure 4 shows another test of the bacteria transformed with both RFP and GFP. The first graph measures GFP fluorescence. (A1, A2, A3, A4, A5 are all clones of the same plasmid with both inducers. ) Figure 5 measures RFP fluorescence. These results show the significant difference in fluorescence between RFP and GFP in the same bacteria; GFP was more expressed compared to the expression of RFP.

While looking back on our experiment, we realized that condensation occurred in the plates that were used in the plate reader which made the Absorbance graph unreliable.


Figure 6

Figure 6 shows the measure of arbitrary fluorescence units of each well in a plate that had only water in its wells, but a lot of condensation on the top of the plate lid. The condensation could therefore have affected our results and resulted in abnormally high levels of absorbance.

Sanger Sequencing Results for GG Parts


Maybe some screenshots of Benchling Successful Results (a list) Unsuccessful Results (a list) The weird, failed backbones that don’t match with anything Parts that we can improve Future Plans The effects of inducers on the fluorescence expression. (IPTG & ATC)