User:Karmella Haynes/Notebook/miR Trigger/Entry Base

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mm/dd/yy Outline of experiments/ tasks Task 1 > Line item 1 Reagent Volume Conc. reagent 1 vol 1 conc 1 reagent 2 vol 2 conc 2 dH2O vol 3     total vol   --> Reaction conditions Lipo transfection > Details Wells Plasmid DNA Volume Lipo Opti-MEM (total) 1 name 2 μg ### μL 4 μL 500 μL 2 name " ### μL " " 3 name " ### μL " " 4 name " ### μL " " 5 name " ### μL " " 6 name " ### μL " " > Add Lipo to 250 μL Opti-MEM --> R.T/ 5 min. > Add DNA to 250 μL Opti-MEM > Add DNA mix to Lipo mix --> R.T./ 20 min. > Remove 500 μL medium from each well; Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37°C > Refresh medium after 5 hours (ab-free) Fugene transfection > Details Wells Plasmid DNA Volume Fugene Opti-MEM 1-6 name 200 ng ### μL 1.8 μL 18.2 μL 7-12 name " ### μL " " 13-18 name " ### μL " " 19-24 name " ### μL " " > Master mixes (4, x6): 10.8 μL Fugene + 109.2 μL Opti-MEM --> R.T/ 5 min. > Add 6x vol DNA --> R.T./ 20 min. > Add 20 μL complexes to each well (1 ml med. each); Grow cells at 37°C overnight