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遗传学
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JoVE 科学教育 遗传学
RNA-Seq
  • 00:00概述
  • 00:59Principles of RNA Sequencing
  • 04:07RNA-Seq Library Preparation and Analysis
  • 07:04Applications
  • 09:06Summary

RNA-Seq

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概述

Among different methods to evaluate gene expression, the high-throughput sequencing of RNA, or RNA-seq. is particularly attractive, as it can be performed and analyzed without relying on prior available genomic information. During RNA-seq, RNA isolated from samples of interest is used to generate a DNA library, which is then amplified and sequenced. Ultimately, RNA-seq can determine which genes are expressed, the levels of their expression, and the presence of any previously unknown transcripts.

Here, JoVE presents the basic principles behind RNA-seq. We then discuss the experimental and analytical steps of a general RNA-seq protocol. Finally, we examine how researchers are currently using RNA-seq, for example, to compare gene expression between different biological samples, or to characterize protein-RNA interactions.

Procedure

RNA sequencing, or RNA-seq, is a technique that can provide information on the sequence and quantity of every RNA expressed, known as the “transcriptome,” in a cell population. Unlike other expression profiling methods such as microarrays, which involve probing for known RNA sequences, RNA-seq can profile gene expression from organisms with un-sequenced genomes. Additionally, RNA-seq can accurately measure a larger range of transcript expression levels than microarrays, especially at very low or very high level…

Disclosures

No conflicts of interest declared.

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JoVE Science Education Database. JoVE Science Education. RNA-Seq. JoVE, Cambridge, MA, (2023).

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