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Diagram:
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Possible ResultsPossible ExplanationControlsProtocolsPartsDone?

YFP visible on surface

success
  • TET-r/BACE2 with Dox
  • TET-r/BACE2 without Dox
  • Hef1a/BACE2
  • No BACE2
PCRPrimers (AL1-4), BACE2 plasmid, eYFP plasmid 
YFP visible in the cellYFP not bound to BACE, YFP disrupts BACE signal sequenceGoldenGate (digest and run on gel to confirm proper assembly)PCR products, GGDonr 
YFP visible out of cellYFP bound to incorrect terminal of BACELR (Gateway) (digest and run on gel to confirm proper assembly)GG product, TRE-t, backbone 
No YFP visibleno Dox, cloning mistakeTransfectLR product, rTTA vector 
no mkate visibletransfection errorAssay Bace2 activity  
  Image  

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