RNA Sequencing

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RNA sequencing (RNA-Seq) is a powerful technique that uses next-generation sequencing (NGS) to reveal the presence and quantity of RNA molecules in a biological sample. Let’s explore the details:

1. Principle of RNA Sequencing:

   • RNA-Seq provides a snapshot of gene expression in a sample, also known as the transcriptome.
   • Unlike gene sequencing (DNA sequencing), which obtains nucleotide sequences of a DNA molecule, RNA-Seq focuses on the order of nucleotide sequences in an RNA molecule.
   • Initially, the Sanger Sequencing technique was used for RNA sequencing, but shortly after the introduction of NGS, high-throughput RNA-Seq became prevalent.

2. Types of RNA Sequencing:

   • Direct RNA Sequencing:
     • In this method, isolated RNAs are sequenced directly without prior conversion into cDNA.
     • RNAs are inherently unstable compared to DNAs, making them challenging to handle.
     • However, direct sequencing avoids biases introduced during cDNA preparation.