ProduceamplifiedamountsofcDNAfromprecious,limitingtotalRNAsampleswithoutintroducingbias.

  • PerformRNAamplificationonpurifiedtotalRNAfollowedbycDNAsynthesisanddetectionFormerly from Epicentre
  • Amplifywiththishigh-fidelity,linearRNAamplificationprocessthatpreservestherelativetranscriptabundanceofthesample.
  • Getmoredataoutofprecioussamples–uselessRNAformoreRT-qPCRreactions
  • Readilyandreproduciblydetectevenlow-abundancetranscriptsinRNAfromasinglecell(CTvalues<35cycles)
  • CollectsmallRNAsampleslessoften.
  • FastprotocolamplifiesandsynthesizescDNAinonly1day.
Applications
  • SignificantlyincreasingthenumberofqRT-PCRsthatcanbeobtainedfromverysmallamountsoftotalRNA,aslittleasonecell(10pg).
  • GenerationoflargeamountsofcDNAfromverysmallsamplesofintacttotalRNAforarchivalpurposes.

TheMessageBOOSTER™cDNASynthesisKitforqPCRenablestheusertoperformsensitiveqPCRamplificationsusingthetotalRNAfromverysmallpopulationsofcells,evenfromaslittleasonecell(Table1).

ThekitamplifiesthemRNAcontainedinasmalltotalRNAsampleandthenconvertstheamplifiedRNAtocDNAthatisreadyforPCRorqPCR(Fig.1).TheMessageBOOSTERcDNASynthesisKitforqPCRusesoligo(dT)toprimecDNAsynthesis.Therefore,thiskitisbestusedwithintacttotalRNApreparations.

Figure1(clicktoenlarge).OverviewoftheMessageBOOSTER™cDNASynthesisKitprocedure.ThekitusesalinearRNAamplificationprocesstoamplifytheRNAfromasmallpopulationofcells,thenconvertstheamplifiedRNAtocDNAthatisreadyforPCR.Theprocedurecanbecompletedin1day.

Figure2.SensitivityoftheMessageBOOSTER™cDNASynthesisKit.cDNAproducedfrom10pgoftotalNRKRNAsignificantlyimprovedthesensitivity(loweredtheCT)ofdetectingthelow-abundancePBGDtranscriptcomparedtocDNAproducedfrom10pgofunamplifiedRNA.

AmountofTotalRNAinaMessageBOOSTER™ReactionNumberofqPCRsThatCanBePerformed
Low-toMedium-AbundanceTranscriptsaMedium-toHigh-AbundanceTranscriptsa
10pg(~1cell)≥10≥100
100pg(~10cells)≥100≥1,000
500pg(~50cells)≥500-1,000≥5,000-10,000
Table1.ThenumberofqPCRamplificationsthatcanbeperformedusingcDNAproducedbyaMessageBOOSTER™cDNASynthesisKit.ThenumberofqPCRsisdependentontheamountofinputtotalRNAandtheabundanceofthetranscript(s)ofinterest.

a.Low-AbundanceTranscripts=1-1,000copiespercell
Medium-AbundanceTranscripts=1,000-10,000copiespercell
High-AbundanceTranscripts=10,000-100,000copiespercell

ProductCitations

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ORDERINFORMATION

MessageBOOSTER™T7-Oligo(dT)PrimerA,RNaseInhibitor,MessageBOOSTER™ReverseTranscriptionPreMix,MessageBOOSTER™DNAPolymerasePreMix1,MessageBOOSTER™DNAPolymerase1,MessageBOOSTER™cDNAFinishingSolution,MessageBOOSTER™RandomPrimers,MessageBOOSTER™RNaseH,MMLVReverseTranscriptase,MessageBOOSTER™InVitroTranscriptionPreMixA,MessageBOOSTER™T7RNAPolymerase,MessageBOOSTER™T7TranscriptionBuffer,RNase-FreeDNaseI,ForwardandReverseControlPCRPrimers,DTT,RNase-FreeWater,Poly(I),NRKTotalRNAControl.