RNA-Seq - Introduction

Introduction

The introduction of next-generation sequencing or high-throughput sequencing technologies opened new doors into the field of DNA sequencing, however as understanding of these technologies becomes more widespread and new tools are developed, new innovative ways of applying these technologies are being created.

Given high-throughput sequencing technologies' low requirements of nucleotide sequence product, together with its deep coverage and base-scale resolution, its use has expanded to the field of transcriptomics. Transcriptomics is an area of research characterizing the RNA transcribed from a particular genome under investigation. Although transcriptomes are more dynamic than genomic DNA, these molecules provide direct access to gene regulation and protein information. Sequencing transcriptomes is not a new idea, however: various methods have been developed previously to directly determine cDNA sequences, based mostly around traditional (and more expensive) Sanger sequencing, while others include methodologies such as Serial analysis of gene expression (SAGE), cap analysis gene expression (CAGE), and massively parallel signature sequencing (MPSS).

Transcriptome Sequencing (RNA-seq) can be done with a variety of platforms to test many ideas and hypotheses. For example, using the Illumina Genome Analyzer platform, recent applications include sequencing mammalian transcriptomes, ABI Solid Sequencing to profile stem cell transcriptomes or Roche's 454 Sequencing to discover SNPs in maize. Even though each platform has its technical differences, the information gathered from each is of the same nature.

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