Team:Duke/Notebook/August
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August 8
Objective: Backbone switching
Delta Ghoshal, Matthew Faw
Redo minipreps from yesterday - culture tubes grew overnight from salvaged cells
- pSB1C3-dCas9-tracrRNA - 171.3 ng/ul
- pSB3C5-R0040-a-tracr 1 - 257.5 ng/ul
- pSB3C5-R0040-a-tracr 2 - 202.7 ng/ul
- pSB3C5-R0040-a-tracr 3 - 200.5 ng/ul
- pSB3C5-R0040-a-tracr 4 - 198.3 ng/ul
Analytical digest of Tube 1 (pSB1C3-dCas9-tracrRNA) using XhoI and BamHI
- 2 hour digest at 37 degrees C
- 3X Master Mix:
- 3 ul Cutsmart
- 22.5 ul water
- 0.75 ul XhoI
- 0.75 ul BamHI
- 9 ul Master Mix added to 1 ul of pdCas9 in one tube and 1 ul of pSB1C3-dCas9-tracrRNA in another tube
- Gel showed BLANK