The why, what, and how of the iso-seq method: using full-length rna ...

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Splice junctions. insufficient connectivity. splice isoform uncertainty ... splice isoform certainty – no assembly required ... -20-hour movie time.

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For Research Use Only. Not for use in diagnostic procedures. © Copyright 2018 by Pacific Biosciences of California, Inc. All rights reserved.The Why, What, and How of the Iso-Seq Method:Using Full-length RNA Sequencing to Annotate Genomes and Solve Diseases Intro to the Iso-Seq MethodJonas Korlach, Chief Scientific Officer WHAT IS THE ISO-SEQ METHOD?The PacBio Iso-Seq method is an end-to-end workflow for sequencing and analyzing full-length transcript isoforms. 1.Convert RNA àcDNA2.cDNA àSMRTbelllibrary 3.Sequence on the Sequel System4.Generate circular consensus sequences (CCS)5.Discover isoforms de novo with Iso-Seq analysis WHY IS FULL-LENGTH RNA SEQUENCING USEFUL?GeneShort-read RNA-seq:Reads spanning splice junctionsInsufficient ConnectivitySplice Isoform UncertaintymRNA isoformsIso-Seq method:Full-length cDNASequence ReadsSplice Isoform Certainty –No Assembly Required Whole-genome Annotation“I would like a reference catalog of all transcript isoforms detectable within a particular...

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