How is RNA sequencing performed and what information details

How is RNA sequencing performed and what information/ details about development it gives? What is the main difference between RNA sequence and microarray when the information is gathered?

Solution

RNA sequencing is reforming the study of transcriptome. This is more responsive and accurate tool for measuring expression across the transcriptome. This provided the visibility to previously undetected changes occurring in disease state. RNA-Seq is used to examine the frequently change in cellular transcriptome. It facilitates the ability to appear in alternative gene spliced transcripts, post-transcriptional modifications, gene fusion, mutations and changes in gene expression, or differences in gene expression in different groups or treatments. In addition to mRNA transcripts, RNA-Seq can look at different populations of RNA to include total RNA or small RNA, such as miRNA, tRNA, and ribosomal profiling. It determines exon/intron boundaries and verify or amend previously annotated 5\' and 3\' gene boundaries.

The development of high-throughput next-generation sequencing (NGS) has revolutionized transcriptomics by enabling RNA analysis through the sequencing of complementary DNA (cDNA). This method has revolutionized our understanding of complex and dynamic nature of the transcriptome. RNA-Seq provides a more detailed and quantitative view of gene expression, alternative splicing, and allele-specific expression. There are many methods for performing an RNA-seq experiment. In fact, the techniques are evolving so rapidly that it can be difficult to decide which one to use. A basic choice is random-primed cDNA synthesis from double-stranded cDNA. Most people will use this method and need to make a further choice between a strand-specific protocol and one that is not.

Difference between RNA-sequence and microarray: RNA sequencing is based on the direct sequencing of cDNA strands by advanced sequencing techniques while Microarray is based on the hybridization potential of predesigned labeled probes with target cDNA sequences. RNA sequencing is performed without the prior knowledge about sequences while Microarray is performed with the prior knowledge about the sequences. Microarray is a robust, reliable and high throughput method used for transcriptome while RNA sequencing is a recently developed whole transcriptome sequencing technique.