The RNA sequencing (RNA-seq) method is a relative abundance measurement technology. The primary goal of the differential gene expression analysis is to quantitatively measure differences in the levels of transcripts between two or more treatments and groups. RNA sequencing (RNA-seq) is based on high-throughput sequencing which will allow a genome-wide detection of transcribed genes. The workflow for RNA-seq is that extracted RNA is converted to cDNA, it is sequenced with a next-generation sequencing platform such as Illumina, and finally, the sequence data are matched to annotated genes by sequence alignment. Data from sequencing will be provided in FASTQ format. Data management includes assessing data for the quality, aligning of the reads to a reference genome, and normalization of the data, before the differential gene expression analysis can be conducted. There are still some technical problems with the technique awaiting resolution, for instance, with respect to PCR amplification bias and bias with the library construction.
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