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</center> | </center> | ||
+ | <h3 class = "text-title"> References </h3> | ||
<center> | <center> | ||
<div class="containertop3"> | <div class="containertop3"> | ||
<div class="item"> | <div class="item"> | ||
− | <img src="https://static.igem.org/mediawiki/2017/ | + | <img src="https://static.igem.org/mediawiki/2017/f/f5/T--TP-CC_San_Diego--finalplasmid.png" height="300"> |
− | <span class="caption"> | + | <span class="caption"> Guide RNA plasmid design </span> |
</div> | </div> | ||
<div class="item"> | <div class="item"> | ||
− | <img src="https://static.igem.org/mediawiki/2017/ | + | <img src="https://static.igem.org/mediawiki/2017/c/c8/T--TP-CC_San_Diego--12WellPlate.png" height="300"> |
− | <span class="caption"> | + | <span class="caption"> 12 Well Plate Design</span> |
</div> | </div> | ||
<div class="item"> | <div class="item"> | ||
− | <img src="https://static.igem.org/mediawiki/2017/ | + | <img src="https://static.igem.org/mediawiki/2017/6/6c/T--TP-CC_San_Diego--100bpLadder.png" height="300"> |
− | <span class="caption"> | + | <span class="caption"> 100 Basepair ladder</span> |
</div> | </div> | ||
</div> | </div> | ||
</center> | </center> | ||
− | |||
− | |||
<h3 class = "text-title">guide RNAs </h3> | <h3 class = "text-title">guide RNAs </h3> | ||
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</center> | </center> | ||
+ | <h3 class = "text-title"> Gels </h3> | ||
<center> | <center> | ||
<div class="containertop3"> | <div class="containertop3"> | ||
<div class="item"> | <div class="item"> | ||
− | <img src="https://static.igem.org/mediawiki/2017/f/f5/T--TP-CC_San_Diego-- | + | <img src="https://static.igem.org/mediawiki/2017/f/f5/T--TP-CC_San_Diego--TLCV2.png" width="400"> |
− | <span class="caption"> | + | <span class="caption"> Digested TLCV2 Vector </span> |
</div> | </div> | ||
<div class="item"> | <div class="item"> | ||
− | <img src="https://static.igem.org/mediawiki/2017/ | + | <img src="https://static.igem.org/mediawiki/2017/8/83/T--TP-CC_San_Diego--pcr.png" width="280"> |
− | <span class="caption"> | + | <span class="caption"> PCR </span> |
+ | </div> | ||
+ | <div class="item"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/0/0f/T--TP-CC_San_Diego--Gel%281%29.jpg" width="350"> | ||
+ | <span class="caption"> GBM39 Genomic DNA </span> | ||
</div> | </div> | ||
</div> | </div> |
Revision as of 03:34, 2 November 2017
Results
Effects of ecDNA
There is a significantly depressed rate of cell growth due to the treatment. New cell growth, measured in the increased of density, was significantly retarded in the induced group. Based on the single factor analysis of variance test, our data after testing declares a p-value deemed low enough in order to reject the null hypothesis.
Parts
Images
EGFR Testing
References
guide RNAs
Gels
Summary
We were able to confirm that the GBM39 cell line had a substantial copy number of the EGFR gene. From there, we designed 4 guide RNAs. Two from exon 1 and two from exon 8. We were unable to design any gRNA using exons 2-7 because in the GBM-39 cell line, that portion of the EGFR gene is mutated. Using the TLCV2 backbone, we removed the stuffer sequence and inserted the guideRNAs in its place. We were successful with constructing 3 gRNAs but the 4th failed to clone twice. After Sequencing our plasmids, we transfected the guideRNAs into GBM39 cells. Currently, we are monitoring the growth of the cells, and confluency and growth of the cancerous cells has been, over the past 2 months, negatively affected in a statistically significant manner - with over a reduction in total confluency growth of over 50% when compared to the control, suggesting that our treatment has an inhibitory effect upon cancer cell growth.