Plate Format:

384 well

Origin:	Tapon Lab
Cell Medium:	Schneiders Drosophila media + 10% heat-inactivated serum + P/S

Protocol:	The ds RNA library plates were thawed for 30mins at RT. Control dsRNA was diluted to 220ng per 5ul in HBSS and then added to the control wells. DNA plasmids UAS-Luc (50ug), G4-DBD-YKi (25ug) and pIZ-Renilla (25ug) were mixed with 21mL EC Buffer and 2.4mL Enhancer. 720ul Effectene transfection reagent was then added and mixed with the S2R+ cell suspension. The cell-DNA complex mix was then dispensed onto the 384well library plates containing the Ambion Library.
Silencing Reagent Concentration:	220ng dsRNA per well
Silencing Reagent Supplier:	Ambion
Silencing Reagent Origin:	Tapon Lab

Cell lysis
Treatment:Aspirate the culture media, add 5ul/well 1x passive lysis buffer
Compounds:passive lysis buffer(1x)
Compound Concentration:5ul/well
Treatment Time:15min
Firefly luciferase activity
Treatment:Add luciferase assay reagent II 5ul/well to read
Compounds:luciferase assay reagent II
Compound Concentration:5ul/well
Treatment Time:Read immediately
Renilla luciferase activity
Treatment:Add Stop&Glo reagent 5ul/well to read
Compounds:Stop & Glo reagent
Compound Concentration:5ul/well
Treatment Time:Read immediately