Next Generation Sequencing. Overview of RNA-seq experimental procedures. Wang L et al. Briefings in Functional Genomics 2010;9:118-128 © The Author.

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Next Generation Sequencing

Overview of RNA-seq experimental procedures. Wang L et al. Briefings in Functional Genomics 2010;9: © The Author Published by Oxford University Press. All rights reserved. For permissions, please

FASTQ format Sequence Quality score encoded in ASCII to save space Machine ID SRA ID & pair info Flowcell laneFlowcell coordinates

Sequence reads aligned using TopHat

Relative expression of transcript (determined by CuffLinks) represented as RPKMs or FPKMs RPKM: Reads Per Kilobase of transcript per Million mapped reads. FPKM: Fragments Per Kilobase of transcript per Million mapped reads.