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- Order LilrB2 and PirB gBlocks
- Simulate Golden Gate on Geneious and make sure it works
- Golden Gate gBlocks into ggDONR
- Transform bacteria with Golden Gate product and plate
- Pick colonies and grow in liquid culture
- Miniprep cells
- Verify miniprep product
- LR into pDEST
- Transform, pick colonies and grow in liquid culture
- Miniprep product
- Midiprep product
- Transfect into HEK293
Status:
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Testing membrane localization of receptor proteins using biotinylated anti-LilrB2 and anti-PirB antibodies.
Plan:
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- Transfect HEK293 with hEF1a:PirB/LilrB2
- Check for membrane localization using anti-PirB/LilrB2 primary antibodies and secondary antibodies conjugated to fluorphores
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Status:
Not yet started. Waiting on Experiment A1.
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