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Related Concept Videos

RNA-seq03:21

RNA-seq

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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. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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Sequencing of mRNA from Whole Blood using Nanopore Sequencing
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Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Published on: June 3, 2019

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Detection of Circulating RNA Using Nanopore Sequencing.

Jennifer Lindemann1, Irene K Yan1, Tushar Patel2

  • 1Mayo Clinic, Jacksonville, FL, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 23, 2021
PubMed
Summary
This summary is machine-generated.

Nanopore sequencing offers comprehensive transcriptome analysis, excelling at profiling long noncoding RNAs. This study details a protocol for using nanopore sequencing of circulating RNA from whole blood for biomarker discovery.

Keywords:
BiofluidsBiomarker discoveryLong noncoding RNANanopore sequencingRNA sequencing

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • RNA sequencing (RNA-seq) is crucial for transcriptome analysis.
  • Long noncoding RNAs (lncRNAs) are important RNA molecules with various functions.
  • Circulating extracellular lncRNAs show potential as disease biomarkers.

Purpose of the Study:

  • To describe a protocol for sequencing circulating RNA using nanopore technology.
  • To enable biomarker discovery from whole blood samples.
  • To leverage nanopore sequencing for comprehensive lncRNA profiling.

Main Methods:

  • Utilized cDNA-PCR amplification of circulating RNA.
  • Employed nanopore sequencing technology for transcript analysis.
  • Applied commercially available kits for sample preparation and sequencing.

Main Results:

  • Demonstrated the feasibility of nanopore sequencing for circulating RNA.
  • Enabled comprehensive profiling of lncRNAs, including novel transcripts and variants.
  • Established a protocol applicable to whole blood samples for biomarker discovery.

Conclusions:

  • Nanopore sequencing provides a powerful and advantageous method for transcriptome analysis, particularly for lncRNAs.
  • The described protocol facilitates biomarker discovery through the analysis of circulating RNA.
  • This approach enhances the potential for identifying novel biomarkers from accessible biological samples.