Optimizedforhighyield,highqualitybiotin-aRNA
HighlyreproducIBLeamplification,labeling,andmicroarrayresults.- MultipleRNAsamplescanbeeasilyamplifiedandlabeledsimultaneously.
- LinearRNAamplificationprocesspreservesthetranscriptprofileofthesample.
- Simple,single-tube,6-hourprocedure.Preparebiotin-aRNAandbeginhybridizationthesameday.
- Biotin-aRNAproducedbythekitdetectsmoretranscripts(genes)thancompetitorkitsandshowedhighcorrelationwithMAQCTaqMan®data(Fig.1).
Applications
TheTargetAmp™1-RoundBiotin-aRNAAmplificationKit105*producesmicrogramamountsofbiotin-antisenseRNA(biotin-aRNA,alsocalledbiotin-cRNA)fromaslittleas25ngoftotalcellularRNA.Thekitprovidesallreagentsforthepreparationofbiotin-aRNA(exceptreversetranscriptaseandRNApurificationcolumns)andusesanimprovedEberwine-typelinearamplificationmethod1toachieveatleast5,000-foldamplificationofthepoly(A)RNApresentinatotalcellularRNAsample.Theentireprocedurecanbecompletedin1day.
Table1.ATargetAmp™Kit105reactionproduceshighyieldsofbiotin-aRNAfromaslittleas25ngofinputtotalRNA.Resultsrepresenttheaverageofmultipleexperiments.
| AmountofInputHeLaTotalRNA | Biotin-aRNAYieldfromHeLaTotalRNAa |
| 25ng | 4.1µg |
| 100ng | 13.7µg |
| 500ng | 64.9µg |
a.Approximately2%ofHeLatotalRNAispoly(A)RNA.
 | Figure1.CorrelationofexpressionratiosobtainedfromqPCRandfrommicroarrayshybridizedtoaRNApreparedwiththeTargetAmp™105kit.Xaxis,log2expressionratiosdeterminedfromHumanGenomeU133Plus2.0Arrays(Affymetrix)hybridizedtotargetspreparedbytheTargetAmp105kit;Yaxis,log2expressionratiosdeterminedbytheMAQC,usingTaqMan®assays.Thehighcorrelationobtained(r=0.936)indicatestheexcellentfidelityoftheamplificationprocess. |
References
- VanGelder,R.N.etal.(1990)Proc.Natl.Acad.Sci.USA87,1663.
*TargetAmp™aRNAAmplificationProductsarecoveredbyintellectualpropertylicensedtoEpicentreTechnologiesCorporationfromJohnson&JohnsonPharmaceuticalResearch&Development,L.L.C.,andbyintellectualpropertysublicensedtoEpicentreTechnologiesCorporationfromIncyteCorporation.
Forresearchuseonly.Someapplicationsinwhichthisproductmaybeusedmaybecoveredbypatentsorpatentapplicationsapplicableincertaincountries.Becausepurchaseofthisproductdoesnotincludealicensetoperformanypatentedapplication,usersofthisproductmayberequiredtoobtainalicense,dependingupontheparticularapplicationandcountryinwhichtheproductisused.
