RNA-Seq

RNA-Seq is a technology that uses the capabilities of next-generation sequencing to reveal a snapshot of RNA presence and quantity from a genome at a given moment in time.

Introduction

RNA-Seq is a revolutionary tool in transcriptomics used for analyzing the continually changing cellular transcriptome. Unlike other methods, RNA-Seq has a far greater resolution and is used to analyze the sum total of RNA transcripts at a given time.

Methodology

RNA-Seq methodology involves the conversion of RNA to cDNA (complementary DNA) using reverse transcription. The cDNA is then fragmented and a library is prepared by ligation of sequencing adapters. The prepared library is then sequenced using next-generation sequencing platforms.

Applications

RNA-Seq has a wide range of applications in biological research including:

• Gene expression profiling
• Detection of alternative splicing
• Identification of novel transcripts
• Detection of gene fusion
• Post-transcriptional modifications

Advantages and Limitations

RNA-Seq has several advantages over other transcriptomics techniques such as its high resolution, low background noise, and the ability to detect structural variants. However, it also has limitations including the need for high-quality RNA and the high cost of sequencing.

See Also

• Next-generation sequencing
• Transcriptomics
• Gene expression
• Alternative splicing

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