www.roche-applied-science.comForlifescienceresearchonly.Notforuseindiagnosticprocedures.0611.108746040017BMCyclinCat.No.10799050001yVersion17Contentversion:June201137.5mg(for2×2.5lmedium)Storeat+2to+8°C1.WhatthisProductDoesKitContentsVial/CapLabelContents1BMCyclin1•25mg,lyophilized•pleuromutilinderivative2BMCyclin2•12.5mg,lyophilized•tetracyclinederivativeStorageandStabilityLyophilizedstableat+2to+8°Cuntiltheexpirationdateprintedonthelabel.NReconstitutedstableat�15to�25°Cforatleast6months.AdditionalEquipmentandReagentsRequiredDAPI*ApplicationBMCyclinisusedfortheeliminationofmycoplasmafrominfectedcellcultureswithoutmarkedcytotoxicsideeffects.SensitiveOrganisms:MycoplasmasandbacteriaOnlyBMCyclinwasfoundtoeffectivelyeliminate•Acholesplasmalaidlawii,•Mycoplasmaarginini,•Mycoplasmahyorhinis,and•Mycoplasmaoralefromexperimentallycontaminatedandchronicallyinfectedcelllines.LThesemycoplasmastrainsaccountformorethan85%ofthecon-taminationsinanimalcellcultures(1).2.HowToUsethisProduct2.1BeforeYouBeginPreparationofStockSolutionDissolvecontentofBMCyclin1andBMCyclin2in10mlsterilePBSorwater.LTheseare250×concentratedstocksolutionsandcanbestoredat�15to�25°Cforatleast6months.RecommendedWorkingConcentrationBMCyclin1:10�g/ml.BMCyclin2:5�g/ml.LTheseconcentrationdonotaffectthegrowthofmostcells.Forsensitivecelllineslowerconcentrationsmaybeused.2.2ProtocolTreatmentofMycoplasma-contaminatedCellCultures�Removeculturemediumfromculturevesselsbyaspiration.�AddnewmediumcontainingBMCyclin1(4�lofstocksolu-tion/ml,finalconcentration10�g/ml).�Cultivatethecellsfor3daysintheusualway�Removeculturemedium.�AddnewculturemediumcontainingBMCyclin2(4�lofstocksolution/ml,finalconcentration5�g/ml).�Cultivatethecellsfor4days.�Repeatthecycletwice.�Checkformycoplasmacontamination(e.g.,withaDNAfluoro-chromesuchasDAPI).2.3DetectionofMycoplasmaswithDAPIAssayPrincipleThefluorescentdyeDAPIbindsselectivelytoDNAandformsstronglyfluorescentDNA-DAPIcomplexeswithhighspecificity.DAPIhasina...
