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Related Concept Videos

siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
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RNA-seq03:21

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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. 
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RACE - Rapid Amplification of cDNA Ends02:35

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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...
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Updated: Dec 8, 2025

A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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Strategies and Best Practice in Cloning Small RNAs.

Hui Dai1, Weifeng Gu1

  • 1Department of Molecular, Cell and Systems Biology, UC Riverside 900 University Ave, Riverside CA 92521, USA.

Gene Technology
|September 21, 2020
PubMed
Summary
This summary is machine-generated.

High-throughput sequencing is crucial for analyzing small RNAs, but library preparation can be challenging. This review compares methods to ensure reliable small RNA sequencing data.

Keywords:
Gene expressionHigh-throughput sequencingRNA librarySmall RNA

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Small RNAs regulate crucial cellular and viral genes.
  • Conventional small RNA analysis methods are often inaccurate and lack sensitivity.
  • High-throughput sequencing offers superior sensitivity and convenience for nucleic acid analysis.

Purpose of the Study:

  • To review and compare strategies for generating high-quality small RNA libraries for high-throughput sequencing.
  • To provide recommendations for best practices in small RNA library preparation.
  • To address challenges in small RNA library construction, including low expression levels and modifications.

Main Methods:

  • Comparative analysis of various small RNA library preparation strategies.
  • Evaluation of methods for overcoming biases in small RNA sequencing.
  • Literature review focusing on recent advancements in high-throughput sequencing library construction.

Main Results:

  • Identification of key challenges in small RNA library preparation, such as low abundance and RNA modifications.
  • Comparison of different library construction protocols regarding their efficiency, accuracy, and reliability.
  • Highlighting strategies that yield representative and unbiased small RNA sequencing libraries.

Conclusions:

  • High-throughput sequencing is essential for comprehensive small RNA analysis.
  • Careful selection of library preparation methods is critical for reliable and accurate results.
  • Adopting best practices ensures the generation of high-quality small RNA libraries for deep sequencing.