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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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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

Published on: August 21, 2019

Cloning new small RNA sequences.

Yuko Tagami1, Naoko Inaba, Yuichiro Watanabe

  • 1Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|March 6, 2010
PubMed
Summary
This summary is machine-generated.

This study presents a method to identify small RNA sequences in plants. The technique purifies small RNAs, ligates adapters, and uses PCR to enable sequencing for analyzing gene regulation and plant responses.

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Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
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Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Published on: November 18, 2014

Area of Science:

  • Plant molecular biology
  • Genomics
  • RNA biology

Background:

  • Small RNAs regulate gene expression and chromatin in eukaryotes.
  • Plant endogenous small RNAs (microRNAs, trans-acting small interfering RNAs) are vital for development and stress.
  • Viral small interfering RNAs (siRNAs) provide antiviral defense via RNA silencing.

Purpose of the Study:

  • To describe a novel method for identifying small RNA sequences from plant tissues.
  • To enable comprehensive analysis of small RNA populations in various plant conditions.

Main Methods:

  • Small RNAs are purified from total RNA using column fractionation and gel excision.
  • Ligation of DNA/RNA chimeric adapters and reverse transcription to cDNA.
  • PCR amplification to add BanI restriction sites for directional concatamerization, followed by cloning and sequencing.

Main Results:

  • The described method allows for the identification of small RNA sequences.
  • The protocol is adaptable for diverse plant samples, including mutants, stressed plants, and virus-infected plants.

Conclusions:

  • This method provides a robust approach for small RNA discovery in plants.
  • It facilitates research into small RNA functions in plant development, stress responses, and viral interactions.