Transcriptomics: Open Access
Open Access

A Short Note on RNA Sequencing

Duhee Bang^* and Byungjin Hwang ^*Correspondence: Duhee Bang, Department of Chemistry, Yonsei University, Seoul, Korea, Email:

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About the Study

RNA-Seq (named as an abbreviation of RNA sequencing) is a sequencing method which usages next-generation sequencing (NGS) to expose the existence and quantity of RNA in a biological sample at a given instant, analyzing the incessantly changing cellular transcriptome. RNA-seq contains change of a sample of RNA to a cDNA library, which is then sequenced and mapped in contradiction of a reference genome. In addition to the facility to measure the level of gene appearance, it offers further info on alternative splicing and non-coding RNA.

RNA sequencing (RNA-Seq) uses the abilities of high-throughput sequencing approaches to provide awareness into the transcriptome of a cell. Compared to preceding Sanger sequencing- and microarray-based approaches, RNA-Seq provides far advanced attention and greater resolve of the dynamic nature of the transcriptome. While sequencing DNA gives a genetic profile of creature, sequencing RNA reproduces only the sequences that are vigorously expressed in the cells. To sequence RNA, the normal method is first to reverse transcribe the RNA removed from the sample to generate cDNA fragments.

There are some sequencing methods and steps used in the RNA

1. mRNA sequencing.

2. Targeted RNA sequencing.

3. Ultra-low-input and single-cell RNA-Seq.

4. RNA exome capture sequencing.

5. Total RNA sequencing.

6. Small RNA sequencing.

7. Ribosome profiling.

A typical RNA-seq experiment consists of the following steps

a. Design Experiment, Set up the experiment to address your questions.

b. RNA Preparation, Isolate and purify input RNA.

c. Prepare Libraries, Convert the RNA to cDNA; add sequencing adapters.

d. Sequence, Sequence cDNAs using a sequencing platform.

e. Analysis.

The mRNA sequence is thus charity as a template to collect in order the chain of amino acids that form a protein. Targeted RNA-sequencing (RNA-Seq) is an extremely precise technique for choosing and sequencing exact transcripts of interest. It offers both measurable and qualitative information. Ultra-low input RNA-Seq has provided a powerful other approach to transcriptomic studies and facilitates new detections with regard to transcriptional dynamics, tissue composition, and regulatory relations between genes. RNA exome capture sequencing overwhelms these experiments by combining RNA-Seq with exome enrichment. This method captures only the coding areas of the transcriptome, letting advanced throughput and needing lower sequencing depth than non-exome capture methods. Whole-transcriptome examination with total RNA sequencing (RNA-Seq) notices coding plus many forms of noncoding RNA. Total RNA-Seq can precisely measure gene and transcript abundance, and classify known and novel features of the transcriptome. Small RNA sequencing is a kind of RNA sequencing created on the use of NGS technologies that lets to isolate and get info about noncoding RNA particles in order to assess and discover new forms of small RNA and to forecast their possible Ribosome profiling, functions. Also known as Ribo-Seq (ribosome sequencing), affords a “snapshot” of all the ribosomes active in a cell at an exact time point. This evidence can help researchers control which proteins are existence actively translated in a cell. Power imitations at different sequencing depths presented that Drop-seq is more cost-efficient for transcriptome quantification of large statistics of cells, while SCRB-seq, MARS-seq, and Smart-seq2 are well-organized when examining fewer cells. The measurable comparison offers the basis for a learned choice among six prominent scRNA-seq procedures, and it offers a framework for benchmarking additional improvements of scRNA-seq protocols.