Anovelhigh-temperatureOneStepRT-PCRkitbasedonthecombinationofSunScript™ReverseTranscriptaseRNaseH-andhigh-qualityTaqPolymeraseforthereversetranscriptionandPCRamplificationofspecificRNAfragmentsinoneconvenientsinglestep.
- ImprovedthermalstABIlityforthereversetranscriptionstep: Worksupto85°C(60-65°Crecommended).
- TranscriptionandamplificationofdifficultRNAs:TranscribesRNAswithhighGCcontentorcomplexsecondarystructures.
- Greatsensitivity: StartswithpicogramamountsofRNA
- ImprovedPCRspecificity:HotstartTaqpolymerasegreatlyimprovesspecificityduringamplification
- HighyieldsofamplifiedDNA
- Highreproducibilityacrossreactions: Bestchoiceforprocessingmultiplesampleswithminimumdeviations.
- Easyhandling: Reagentsareaddedinasinglestep,whichalsominimizesdangersofcontaminationandreducesoverallhandlingtime.
TheSunScript™OneStepRT-PCRKitisaneasyandreliablesystemdesignedforfast,specificandsensitiveendpointRT-PCRreactions.ThiskitcontainsallthecomponentsneededtoperformbothreversetranscriptionandPCRamplificationinasingletubebyusinggenespecificprimersina“onestep”reaction.Thisminimizescontaminationsandconstitutesaconvenientsystemtoprocessmultiplesamples.
- OneStepRT-PCRProtocolOverview
- DetectionofmRNAswithdifferentexpressionlevels
- HighthermalactivityoftheRTstep
- Highsensitivity
- Highreproducibility
- AmplificationanddetectionofdifficultRNAtargets
OneStepRT-PCRProtocolOverview
Backtotop
DetectionofmRNAswithdifferentexpressionlevels
100ngoftotalRNAfromHeLacellswerereversetranscribedandamplifiedwithSunScript™OneStepRT-PCRKitandspecificprimersfortheindicatedgenes(GAPDH,HERCGUSB,CLTCandACTIN).
Thekiteasilydetectsgeneswithvastlydifferentexpressionlevels.
Backtotop
HighthermalactivityoftheRTstep
100ngoftotalRNAfromHeLacellswerereversetranscribedandamplifiedwithSunScript™OneStepRT-PCRKitandspecificprimersfortheGAPDHopenreADIngframe(productlength305bp).
TheRTreactioncanbeperformedattemperaturesupto85°C,makingthisthebestchoicefordetectingdifficultRNAs,withhighdegreesofsecondarystructure.
Backtotop
Highsensitivity
TheindicatedamountsoftotalRNAfromHeLacellswerereversetranscribedandamplifiedwithSunScript™OneStepRT-PCRKitandspecificprimersforGAPDHCDNA(productlength305bp).
ThesensitivityofthekitisgoodenoughtodetectexpressedgenesfromtotalRNAstartingmaterialintheorderofpicograms.
Backtotop
Highreproducibility
100ngoftotalRNAfromHeLacellswerereversetranscribedandamplifiedwithSunScript™OneStepRT-PCRKitandspecificprimersfortheindicatedgenes(4replicatesareperformedpergene).
Thefourindependentreplicatespergeneshowsimilarsensitivityandyield,makingthiskitthebestchoiceforprocessingmultiplesampleswithminimumdeviations.
Backtotop
AmplificationanddetectionofdifficultRNAtargets
100ngoftotalRNAfromHeLacellswerereversetranscribedandamplifiedwithSunScript™RT-PCRKitandfourdifferentspecificprimerpairsforafragmentoftheTAFII100cDNAwith75%GCcontent(productlengths430-470bp).
TheRT-PCRprotocolwithsampledenaturationwasperformedhereandthecDNAsynthesisstepwasperformedat60°C.
ThefouroligopairsproducedeffectiveamplificationbandsdespitetheelevatedGCcontentofthetemplateRNA.
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ORDERINFORMATION
EachSunScript™OneStepRT-PCRKitcontains:EnzymeMix,2XReactionBufferandwater.ThecompletemanualisavailableundertheManualtab.LucigenisanauthorizeddistributorofSygnisproductsintheUS.
SunScript™OneStepRT-PCRKitisintendedformolecularBIOLOGyuseonlyandinvitrouseonly.Thisproductisnotintendedfordiagnosis,preventionortreatmentofadiseaseinhumanbeingsoranimals.
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