Difference between revisions of "Team:TP-CC San Diego/Results"

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         <span class="caption"> EGFR vs. Normalized Vector Expansion</span>
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         <span class="caption"> E1G4</span>
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         <span class="caption"> EGFR vs. Normalized Vector Expansion</span>
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         <span class="caption"> GBM39 Genomic DNA </span>
 
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Revision as of 21:48, 30 October 2017

Results

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.

Images

STBL3 competent cell colonies containing guide RNAs

E1G2
E1G4
E8G2
E8G3

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

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