1. 6-WellPlates
   1. Bathing
      1. PreparedsRNAsUSPendedinwater.Weuse~500bpdsRNA.
      2. Add~10-30µgdsRNAtowellsof6-welltissuecultureplate.Weuse0.1-0.3µgin384-wellplatesfor25-50nMfinalconcentration.
      3. Countcells,thenspintopellet(~1200rpm,5").
      4. Resuspendcellsat1-5x106cells/mlinserumfreemedia.
      5. Plate1mlcellsintowellsof6-wellplate.Itdoesn"tseemtomatterifdsRNAorcellsareaddedfirst.
      6. IncubatedsRNAwithcellsatRTfor30".
      7. Add3mlcompletemediawith10%FBStoeachwell.
      8. Incubate3daysandanalyze.Lengthofincubationmayvarydependingonassay.
2. 384-WellPlates
   1. Bathing
      1. Remove384-wellplatespre-aliquotedwithdsRNAfromfreezertothaw.The384-wellplatescontain5ulof~0.05ug/uldsRNAinwaterfor~0.25ugdsRNA/well.ThedsRNAsare~500bp.
      2. Spinplatesat~1200rpmfor1".beforeremovingseals.
      3. Countcells,thenspintopellet(~1200rpm,5").
      4. Resuspendcellsat1-5x106cells/mlinserumfreemedia.
      5. Plate10ulcellsintowellsof384-wellplate.
      6. IncubatedsRNAwithcellsatRTfor30".
      7. Add30ulofcompletemediatoeachwell.
      8. Sealtheplatestopreventevaporation.
      9. Incubate3daysandanalyze.Lengthofincubationmayvarydependingonassay.
   2. Transfection
      1. Remove384-wellplatesfromfreezertothaw.The384-wellplatescontain5ulof~0.016ug/uldsRNAinwaterfor~0.08ugdsRNA/well.ThedsRNAsare~500bp.
      2. Spinplatesat~1200rpmfor1".beforeremovingseals.

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