ProductDescription

First-strandCDNASynthesisKit25ReactionKit.

CONTENTS:

Thiskitcontainssufficientreagentstoperform25firststrandcDNAsyntheseswithupto2mgofpurifiedpolyA(+)mRNAor10mgoftotalcellRNAperreaction.

STORAGE:

12monthsat-20°C
12monthsat-70°C

DESCRIPTION:

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.

cDNASYNTHESIS

Figure1.[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.

FORLABORATORYUSEONLY

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