首页 > Lifesci > Lifesci/DirectScript cDNA Synthesis Kit/LSK1200-25/25 reactions

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Lifesci/DirectScript cDNA Synthesis Kit/LSK1200-25/25 reactions

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LSK1200-25

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Lifesci

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4000-520-616

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ProductDescriptionFirst-strandCDNASynthesisKit25ReactionKit.CONTENTS:Thiskitcontainssufficientreagentstoperform25firststrandcDNAsyntheseswithupto2mgofpurifiedpolyA(+)mRNAor10mgoftotalcellRNAperreaction.STORAGE:12monthsat-20°C12monthsat-70°CDESCRIPTION:ThisFirst-strandcDNASynthesiskithasbeendevelopedtomaximizethesynthesisoffull-lengthcDNA.ThiskitproducesanoptimaltemplateforDNAamplificationtechniques.Testedagainsta7.5kbsyntheticpolyA(+)RNAanda9.8kbnativeviralmRNA,32%and42%ofthecDNAssynthesizedwerefull-length,respectively.Also,cDNAstoapolyA(+)RNAladderconsistedofapproximately70%fulllengthproduct(Fig.1).Theuseoffull-lengthcDNAtemplatesforpolymerasechainreaction(PCR)(1)affordsequalavailABIlityofallsequencesforprimingandamplificationregardlessoflocation.ThiskitincludesRNaseinhibitor,DTTsolution,5Xreactionbuffercontainingnucleotides,andAMVreversetranscriptaseenzyme.Pd(T)12-18andrandomhexamerprimersarealsoprovided,togetherwithRNase-freewaterandapolyadenylatedRNAladderascontroltemplate.cDNASYNTHESISFigure1.[32P]-labeledcDNAtothreeRNAtemplates:Lanes1and2:7.5kbsyntheticpolyA(+)RNAprimedwithpd(T)12-18andrandomhexamers,respectively.Lanes3and4:9.8kbtobaccoveinmottlingvirus(TVMV)mRNA,primedwithpd(T)12-18andrandomhexamers,respectively.Lanes5and6:polyA(+)RNAladder*primedwithpd(T)12-18andrandomhexamers,respectively.*ThispolyA(+)RNAladderisdifferentfromtheonesuppliedinthiskit.FORLABORATORYUSEONLYDownloaddetailsandprotocol.