- Purchase BACE2 plasmid (http://dnasu.org/DNASU/GetCloneDetail.do?cloneid=76140)
- Design primers for BACE2 and EYFP
- Run PCR on BACE2 and EYFP
- Perform golden gate on PCR products and GGDonr
- Perform LR reaction with products from previous step, TRE-t, and backbone
- Transfect with expression vector from previous step and rtta vector as well as HEF1a + mkate
- Digest portion with Bal1, run gel and compare to virtual gel
-
Observe for fluorescence in presence of Dox (more work needed on details of actual experiment)
...