Determining the sequence of RNA. This technique is used for discovery and quantification of mRNAs and ncRNAs.
Involves library preparation and sequencing. Library preparation might slightly vary for different classes of RNAs. The general procedure for mRNA and most ncRNA sequencing involves:
- Enrichment of polyA RNAs using oligo-dT beads to remove ribosomal RNA. Alternatively, ribosomal RNAs can be selectively removed. Refer to this.
- cDNA preparation
- Adapter ligation
- PCR amplification
See also dna-sequencing.