Total RNA sequencing /Whole transcriptome sequencing refers to sequencing technology used to determine the sequences of RNA molecules for characterizing the RNA content and composition of a given sample. It includes mRNAs, non-coding RNAs and small RNAs. It is a genetic tool for understanding the transciptome, which in-turn is essential for interpreting the functional regions of the genome and unfolding the molecular constituents of cell and tissues, and also for understanding the relative change of transcript during defined developmental stages or under specific condition.
1. Quantification and characterization of transcript of an organism
2. Identification of known and novel transcripts, isoforms and gene fusion
3. Splice junction analysis
4. SNP analysis: Identification of SNPs in the transcribed region which can be used as biomarkers for profiling large population.
5. Differential gene expression profiling
6. de novo assembly and annotation
1. RNA Extraction
2. RNA quality check using Qubit and Bioanalyser
3. Preparation of the sequencing libraries
4. Validation of the prepared library
5. Sequencing (HiSeq/NextSeq/Pacbio)
6. Bioinformatics Analysis