mRNA sequencing adopts next-generation sequencing technique (NGS) to capture the messenger RNA(mRNA) form Eukaryote at specific period that some special functions are activating. The longest transcript spliced was called ‘Unigene’ and used as the reference sequence for subsequent analysis, which is an effective means to study the molecular mechanism and regulatory network of the species without reference.
After transcriptome data assembly and unigene functional annotation
(1)SNP analysis, SSR analysis, CDS prediction and gene structure will be preformed.
(2)Quantification of unigene expression in each sample will be performed.
(3)Differentially expressed unigenes between samples (or groups) will be discovered based on unigene expression
(4)Clustering, functional annotation and enrichment analysis of differentially expressed unigenes will be performed