Diagram:
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| Possible Results | Possible Explanation | Controls | Protocols | Parts | Done? |
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YFP visible on surface | success |
| PCR | Primers (AL1-4), BACE2 plasmid, eYFP plasmid | |
| YFP visible in the cell | YFP not bound to BACE, YFP disrupts BACE signal sequence | GoldenGate (digest and run on gel to confirm proper assembly) | PCR products, GGDonr | ||
| YFP visible out of cell | YFP bound to incorrect terminal of BACE | LR (Gateway) (digest and run on gel to confirm proper assembly) | GG product, TRE-t, backbone | ||
| No YFP visible | no Dox, cloning mistake | Transfect | LR product, rTTA vector | ||
| no mkate visible | transfection error | Assay Bace2 activity | |||
| Image |
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Nanodrop from YFP PCR products with original primers :
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239 299 ng/uL |