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Whole transcriptome analysis with sequencing: methods, challenges and potential solutions

Jiang, Zhihua, Zhou, Xiang, Li, Rui, Michal, Jennifer J., Zhang, Shuwen, Dodson, Michael V., Zhang, Zhiwu, Harland, Richard M.
Cellular and molecular life sciences 2015 v.72 no.18 pp. 3425-3439
biomarkers, gene expression, genes, high-throughput nucleotide sequencing, messenger RNA, pathogens, polymerase chain reaction, transcriptome, transcriptomics
Whole transcriptome analysis plays an essential role in deciphering genome structure and function, identifying genetic networks underlying cellular, physiological, biochemical and biological systems and establishing molecular biomarkers that respond to diseases, pathogens and environmental challenges. Here, we review transcriptome analysis methods and technologies that have been used to conduct whole transcriptome shotgun sequencing or whole transcriptome tag/target sequencing analyses. We focus on how adaptors/linkers are added to both 5′ and 3′ ends of mRNA molecules for cloning or PCR amplification before sequencing. Challenges and potential solutions are also discussed. In brief, next generation sequencing platforms have accelerated releases of the large amounts of gene expression data. It is now time for the genome research community to assemble whole transcriptomes of all species and collect signature targets for each gene/transcript, and thus use known genes/transcripts to determine known transcriptomes directly in the near future.