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Related Concept Videos

RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
Since the...

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Related Experiment Video

Updated: Jun 9, 2026

Preparation of Small RNA Libraries for Sequencing from Early Mouse Embryos
08:37

Preparation of Small RNA Libraries for Sequencing from Early Mouse Embryos

Published on: October 9, 2020

Construction of small RNA cDNA libraries for deep sequencing.

Molly F Thomas1, K Mark Ansel

  • 1Department of Microbiology & Immunology, Strategic Asthma Basic Research Center, University of California San Francisco, San Francisco, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 10, 2010
PubMed
Summary

Small RNAs, particularly microRNAs (miRNAs), regulate mammalian immune cell development. Next-generation sequencing offers a powerful method for profiling small RNA transcriptomes and discovering novel small RNAs.

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Last Updated: Jun 9, 2026

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Area of Science:

  • Molecular Biology
  • Genetics
  • Immunology

Background:

  • Small RNA-mediated gene silencing is crucial for cell lineage establishment.
  • MicroRNAs (miRNAs) play significant roles in mammalian immune system development and function.
  • Next-generation sequencing (NGS) is a key technology for small RNA transcriptome profiling.

Purpose of the Study:

  • To highlight the importance of small RNAs, especially miRNAs, in immune cell development.
  • To emphasize the utility of NGS for comprehensive small RNA analysis.
  • To discuss the methods for generating small RNA libraries for sequencing.

Main Methods:

  • Small RNA library preparation involving adapter ligation, reverse transcription, and PCR amplification.
  • High-throughput sequencing using next-generation sequencers for parallel processing of millions of reads.
  • Comparison of NGS with traditional Sanger sequencing and hybridization-based techniques.

Main Results:

  • NGS enables rapid generation of large datasets (megabases) from diverse cell types.
  • NGS is superior to other methods for discovering novel small RNAs and identifying posttranscriptional editing.
  • The small RNA transcriptome can be effectively profiled using NGS.

Conclusions:

  • Small RNAs, particularly miRNAs, are critical regulators of immune cell biology.
  • Next-generation sequencing is an indispensable tool for advancing small RNA research.
  • High-throughput sequencing facilitates the discovery of novel small RNAs and regulatory mechanisms.