The Daily Grind
- Doing ACC deaminase assay on PSBA::ACCD::GfpUV in pDSK-GFPuv as per "Penrose and Glick" -- differences are that I incubated the bacteria in TSB at 37 degrees overnight, resuspended them in 4 mL of M9 minimal media without N and added 50 ul of unfiltered 0.5 M ACC in water. They are now shaking at 25 degrees until tommorow.
- Finished the protocol the next day, and both cultures absorbed 0.174 A at 540 nm. Translation, the ACC deaminase is non-functional which corroborates the lack of growth on nitrogen free media. Will retry amplifying both again and re-cloning with low copy promoters.