RNAszekvenálást

RNA sequencing, often abbreviated as RNA-Seq, is a powerful molecular biology technique used to determine the presence and quantity of RNA in a biological sample. It utilizes next-generation sequencing (NGS) technologies to sequence complementary DNA (cDNA) molecules that are reverse-transcribed from RNA. This process allows for the comprehensive analysis of the transcriptome, which is the complete set of RNA transcripts produced by an organism or cell at a specific time.

The workflow typically begins with the isolation of total RNA or specific RNA fractions from a sample. This RNA is then converted into cDNA using an enzyme called reverse transcriptase. Next, cDNA fragments are prepared for sequencing, which involves processes like fragmentation, adapter ligation, and amplification. The prepared library is then sequenced on an NGS platform, generating millions of short sequence reads. These reads are subsequently aligned to a reference genome or transcriptome, or assembled de novo if a reference is unavailable.

The primary applications of RNA-Seq include quantifying gene expression levels, identifying novel transcripts and splice variants, discovering gene fusions, and analyzing post-transcriptional modifications. By comparing RNA-Seq data from different conditions, researchers can gain insights into biological processes, identify disease biomarkers, and understand cellular responses to various stimuli. Its high sensitivity and dynamic range make it a preferred method for transcriptomic studies.