TransfectionReagent(invivo)ishighlyeffectiveforDNAplasmiddeliveryinvivo,whilenotinducinganinflammatoryresponse.TurboFectinvivohasbeenassessedforplasmidDNAdeliveryinbothmiceandratsandissuitableforDNAadmiNISTrationviavariousroutes,includingintravenous,intraperitoneal,andintratumoralinjection.TheamountofDNAandsuggestedinjectionvolumewilldependontheexperimentalanimalandtherouteofadministrationaswellasthetargetedtissueororganandtheexpressionvector.||TransfectionReagent(invivo)isasterilesolutionofacationicpolymerinwater.Thepolymerformscompact,stable,positivelychargedcomplexeswithDNA.ThesecomplexesprotectDNAfromdegradationandfacilitategenedeliveryinvivo.ReagentcausesnodetectableinflamatoryresponseandissuitableforDNAadministrationviavariousroutes,includingintravenous,intraperitoneal,intratumoralinjection,etc.||TransfectionReagent(invivo)for20transfectionsusing50ugDNA.||ReagentsRequiredbutnotSupplied:|Sterile5%glucosesolutionfordilutionofDNA.||GeneralConsiderations:||DNAQualityRequirements:|DNAqualityiscriticalforsuccessfultransfection.AnA260/A280ratioof1.8orhigherisrecommended.DNAshouldbesterileandfreeofanycontaminantssuchasendotoxins.||ChoiceofPromoter:|Highgeneexpressiondependsonboththepromoterunderwhichthegeneofinterestisexpressedandthetargetedtissue/organ.Cytomegalovirus(CMV)promoterisbestknownforhighgeneexpressioninawidevarietyofcells.SomeresearcherspreferLTR,simianvirus(SV40)orRoussarcomavirus(RSV)promoters.||TransfectionReagent/DNARatio:|ThevolumeoftransfectionreagentuseddependsontheamountofDNA,transgeneandanimalstobetransfected.Theratiospresentedintheprotocolsbelowarestartingamountsandcanbefurtheroptimizedforbestresults.||AmountofDNAandsuggestedinjectionvolume:|TheamountofDNAandsuggestedinjectionvolumedependontheexperimentalanimalandtherouteofadministration(seeprotocol)aswellasthetargetedtissueororganandtheexpressionvector.TopreventprecipitationofTransfectionReagent/DNAcomplexes,thefinalconcentrationofDNAintheinjectionmixshouldnotexceed0.5ug/ul.||QualityControl:|TransfectionefficiencywastestedonBALB/cfemalemice(7-8weeksold)using50ugFireflyluciferasecodingplasmidand6ulofinvivoTransfectionreagentspermouse.ThedeliveryofthetransgenewasevaluatedbymeasurementofFireflyluciferaseactivityintotalproteinextractedfromthelungsoftransfectedanimals.100%oftestedmiceexpressedtheFireflyluciferase.||StorageandStABIlity:|Maybestoredat4°C.Forlong-termstorage,aliquotandstoreat4°C.Donotfreeze.Aliquotsarestablefor6monthsafterreceipt.Formaximumrecoveryofproduct,centrifugetheoriginalvialpriortoremovingthecap.

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