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RNA Captor A Tool for RNA Characterization 英文参考文献
RNACaptor:AToolforRNACharacterization
ChristianClepet*
URGVPlantGenomics,INRAUMR1165UEVE/CNRSERL8196,Evry,France
Abstract
Background:Inthegenomeera,characterizingthestructureandthefunctionofRNAmoleculesremainsamajorchallenge.
Alternativetranscriptsandnon-protein-codinggenesarepoorlyrecognizedbythecurrentgenome-annotationalgorithms
andefficienttoolsareneededtoisolatetheless-abundantorstableRNAs.
Results:AuniversalRNA-taggingmethodusingtheT4RNAligase2andspecialadaptersisreported.Basedonthissystem,
protocolsforRACEPCRandfull-lengthcDNAlibraryconstructionhavebeendeveloped.TheRNAtaggingconditionswere
thoroughlyoptimizedandcomparedtopreviousmethodsbyusingabiochemicaloligonucleotidetaggingassayandRACE
PCRs on a range of transcripts. In addition, two large-scale full-length cDNA inventories relying on this method are
presented.
Conclusion:TheRNACaptorisastraightforwardandaccessibleprotocol.Thesensitivityofthisapproachwasshowntobe
highercomparedtopreviousmethods,andapplicableon messengerRNAs,non-protein-coding RNAs,transcription-start
sitesandmicroRNA-directedcleavagesitesoftranscripts.ThisstrategycouldalsobeusedtostudyotherclassesofRNAand
indeepsequencingexperiments.
Citation:ClepetC(2011)RNACaptor:AToolforRNACharacterization.PLoSONE6(4):e18445.doi:10.1371/journal.pone.0018445
Editor:JuanMata,UniversityofCambridge,UnitedKingdom
ReceivedJanuary3,2011;AcceptedFebruary28,2011;PublishedApril13,2011
Copyright: ?2011 Christian Clepet. This is anopen-access article distributed under the terms of the Creative Commons Attribution License, whichpermits
unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited.
Funding:ThisworkwassupportedbyUMR1165INRAUEVE,CNRSERL8196,theInternationalGrapevineGenomeProjectANR-05-GPLA-014andtheInternational
CucurbitGenomeInitiative.Thefundershadnoroleinstudydesign,datacollectionandanalysis,decisiontopublishorpreparationofthemanuscript.
CompetingInterests:TheauthorwasgrantedapatentonthisRNA-taggingmethod
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