Informatics for RNA Sequencing: A Web Resource for Analysis on the Cloud
Fig 3
RNA-seq library fragmentation and size selection strategies that influence interpretation and analysis.
RNA-seq library construction may involve both fragmentation and size selection. These procedures may be modified according to the integrity and amount of starting total RNA. The distributions of RNA molecule sizes are depicted for input total RNA and at various stages during the process of RNA/cDNA fragmentation and size selection. Commonly used methods for fragmentation and size selection are depicted along with the expected output of a quality-control assay at each stage (in the form of a capillary electrophoresis trace). Note that in the final library, it is typical that the majority of RNAs below a certain size (typically <150–200 bp) are underrepresented. Refer to S3 Table and S7 Table for more details on many of the concepts depicted in this figure.