User:Karmella Haynes/Notebook/miR Trigger/2010/05/06

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05/06/10

  • ✓ CFP reporter lines: FC32 and FC34 in JDS33-47



CFP reporter lines
> Continuing Francesca's work
> Goal: make stables via random insertion (single copy may be too low).
> Challenge: CFP reporters are puro-marked, but JDS line is already puro resistant


> Colony plates:
--> No CFP signal in any of the large plates
--> Very few (< 1%) CFP+ cells in back-up non-clonal plates (6-well, 3 wells per "line")
--> Cells grow just as well in 2.0 μg/mL puro as in 1.0 μg/mL puro


> Today, using 6-well back-up cultures (3 wells per "line"):
--> For each "line", plate 50% of 1 well in 2.0 μg/mL puro med. and 50% in 5.0 μg/mL puro med.
--> For each line, split another well over 3 new wells in 6-well plate