Next Gen Sequencing

RNA-Seq

Journal.pcbi.1004393.g002

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.
RNA-Seq-alignment

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:

RNASeqWorkflow2016

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