Guidelines for RNA-seq projects: applications and opportunities in non-model decapod crustacean species

Next-generation sequencing (NGS) has dramatically changed the way biological research is being conducted in the post-genomic era, and they have only been utilized widely over the recent decade for studies of non-model decapod crustacean species, predominantly by sequencing the transcriptome of various tissues across different life stages. Next-generation sequencing can now provide a rapid, cost-effective solution for discovery of genetic markers crucial in many applications that would previously have otherwise taken years to develop. Sequencing of the entire transcriptome (referred to as RNA sequencing; RNA-seq) is one of the most popular NGS tools. RNA-seq studies of non-model species in crustacean taxa, however, have faced some problems, including a lack of “good” experimental study design, a relative paucity of gene annotations, combined with limited knowledge of genomic technologies and analyses. The aim of the current review is to assist crustacean biologists to develop a better appreciation for the applications and scope of RNA-seq analysis, understand the basic requirements for optimal RNA-seq studies and provide an overview of each step, from RNA-seq experimental design to bioinformatics approaches to data analysis. Insights that have resulted from RNA-seq studies across a wide range of non-model decapod species are also summarized.