Selective Depletion of rRNA Enables Whole Transcriptome Profiling of Archival Fixed Tissue 英文参考文献.docVIP

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Selective Depletion of rRNA Enables Whole Transcriptome Profiling of Archival Fixed Tissue 英文参考文献.doc

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Selective Depletion of rRNA Enables Whole Transcriptome Profiling of Archival Fixed Tissue 英文参考文献

SelectiveDepletionofrRNAEnablesWhole TranscriptomeProfilingofArchivalFixedTissue JohnD.Morlan*,KunbinQu,DominickV.Sinicropi GenomicHealthInc.,RedwoodCity,California,UnitedStatesofAmerica Abstract We report a method for Selective Depletion of abundant RNA (SDRNA) species from Human total RNA isolated from formalin-fixed, paraffin-embedded (FFPE) tissue, here demonstrating removal of ribosomal and mitochondrial transcripts fromclinicalFFPEtissueRNAarchivedupto20years.Importantly,SDRNAremoves98%oftargetedRNAswhilepreserving relative abundance profiles of non-targeted RNAs, enabling routine whole transcriptome analysis of clinically valuable archivaltissuespecimensbyNext-GenerationSequencing. Citation:MorlanJD,QuK,SinicropiDV(2012)SelectiveDepletionofrRNAEnablesWholeTranscriptomeProfilingofArchivalFixedTissue.PLoSONE7(8):e42882. doi:10.1371/journal.pone.0042882 Editor:SoheilS.Dadras,UniversityofConnecticutHealthCenter,UnitedStatesofAmerica ReceivedMarch22,2012;AcceptedJuly13,2012;PublishedAugust10,2012 Copyright: ? 2012 Morlan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:Nocurrentexternalfundingsourcesforthisstudy. CompetingInterests:TheauthorsaresalariedemployeesofGenomicHealth,Inc.andholdstockofthecompany.Apatentapplicationrelatingtothemethod describedinthismanuscripthasbeenfiled,U.S.ApplicationNo.12/940,981.Thisdoesnotaltertheauthors’adherencetoallPLoSONEpoliciesonsharingdata andmaterials. *E-mail:jmorlan@ Introduction about an hour and the resulting depleted RNA can be used as inputtovirtuallyanycDNAlibraryconstructionmethod. ThelargerepositoriesofFFPEarchivaltumorspecimenslinked tolongtermclinicalrecordsprovidearichresourcefordiscovery of prognostic and predictive biomarkers. Whole-transcriptome analysis by Next-Generation Sequencing (NGS) technologies provides an opportunit