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Team:Cambridge/Notebook/Week5
From 2010.igem.org
Monday
We took photographs of the petri dishes that Theo and Peter had drawn. These had grown up well, over a short period. We believe this is because they were plated out with only ampicillin, and no chloramphenicol.
Will designed oligos to be sent to Biolegio for synthesis. Hannah and Emily wrote more messages on dishes, and planned a restriction digest to check for the biobrick compatibility of the V. fischeri lux operon.
Anja and Theo miniprepped an overnight culture which had been inoculated with a ligation of an expression backbone and firefly luciferase. They ran it on a gel, which showed that the insert had worked. Discovered that E-gels can be used multiple times. Then they performed a very bodged luciferase assay. 0.5ml of cells were frozen and thawed three times to lyse them, then 0.05ml of a concentrated D-luciferin solution was added. The camera showed them to be glowing.
Tuesday
Will constructed oligos, and taught Emily and Hannah to do so. V. phosphoreum arrived, Anja and Theo made up its specialised, salty culture. Peter and Theo placed it in five subcultures and inoculated.
Wednesday
Thursday
Friday
