RNASeq library preparation

Total RNA

mRNA-seq workflow

Illumina NGS Library

Illumina library fragment

Illumina library read bindings 1

Illumina library read bindings 2

RNA-Seq applications

• rRNA depleted samples
• Very high dynamic range
• No prior knowledge of expressed genes.

- Gives more information than microarrays.

• Applications

- Sequencing of mRNA

- Differential expression of known or unknown transcripts during a treatment or condition

- Isoform detection.

- Alternative splicing events

- Post-transcriptional mutations or editing

- Gene fusions

- Non-coding RNAs

- miRNAs/small non-Coding RNA

NGS Data Analysis

The number of replicates issue

• http://rnajournal.cshlp.org/content/22/6/839.short

• http://www.biorxiv.org/content/early/2016/12/02/090753