Next Gen Sequencing


Credit: Malachi Griffith , Jason R. Walker, Nicholas C. Spies, Benjamin J. Ainscough, Obi L. Griffith (CC-BY 4.0)
Credit: Rgocs (CC-BY)
RT-PCR and RT-qPCR can be used to measure the abundance of specific transcripts in a fairly low throughput way. Leveraging the the concept of Reverse Transcription and coupling that to high-throughput sequencing technologies, transcripts can be sequenced and mapped to a genome to depict the quantity of transcripts as represented by number of reads.

Given sufficient read coverage, novel splice isoforms can also be identified as different exon-exon junctions are identified.


The general workflow of RNA-Seq analysis follows:

Credit: Salubrious Toxin (CC-BY-SA 4.0)