Difference between revisions of "Team:TP-CC San Diego/Results"
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<h3 class ="day">Parts</h3> | <h3 class ="day">Parts</h3> | ||
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<img src="https://static.igem.org/mediawiki/2017/d/d5/T--TP-CC_San_Diego--k2369000.png"> | <img src="https://static.igem.org/mediawiki/2017/d/d5/T--TP-CC_San_Diego--k2369000.png"> | ||
<span class="caption">BBa_K2369000</span> | <span class="caption">BBa_K2369000</span> | ||
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<img src="https://static.igem.org/mediawiki/2017/7/7b/T--TP-CC_San_Diego--k2369001.png"> | <img src="https://static.igem.org/mediawiki/2017/7/7b/T--TP-CC_San_Diego--k2369001.png"> | ||
<span class="caption">BBa_K2369001</span> | <span class="caption">BBa_K2369001</span> | ||
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<img src="https://static.igem.org/mediawiki/2017/f/f7/T--TP-CC_San_Diego--k2369002.png"> | <img src="https://static.igem.org/mediawiki/2017/f/f7/T--TP-CC_San_Diego--k2369002.png"> | ||
<span class="caption">BBa_K2369002</span> | <span class="caption">BBa_K2369002</span> | ||
Revision as of 01:02, 2 November 2017
Results
Parts
BBa_K2369000
BBa_K2369001
BBa_K2369002
Images
EGFR copy number
EGFR vs. Normalized Vector Expansion
E1G4
Gels
Digested TLCV2 Vector
PCR
GBM39 Genomic DNA
References
Guide RNA plasmid design
12 Well Plate Design
100 Basepair ladder
STBL3 competent cell colonies containing guide RNAs
E1G2
E1G4
E8G2
E8G3
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.
