Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: Oct 11, 2025

Sequencing of mRNA from Whole Blood using Nanopore Sequencing
11:26

Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Published on: June 3, 2019

14.0K

Nanopore Whole Transcriptome Analysis and Pathogen Surveillance by a Novel Solid-Phase Catalysis Approach.

Yi Fang1, Amogh Changavi1, Manyun Yang2

  • 1New England Biolabs, Inc., Ipswich, MA, 01938, USA.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|November 27, 2021
PubMed
Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ginkgo biloba Extract Attenuates the Disruption of Pro- and Anti-inflammatory Balance of Peripheral Blood in Arsenism Patients by Decreasing Hypermethylation of the Foxp3 Promoter Region.

Biological trace element research·2022
Same author

In utero exposure to methylmercury impairs cognitive function in adult offspring: Insights from proteomic modulation.

Ecotoxicology and environmental safety·2022
Same author

LC/MS/MS-Based Liver Metabolomics to Identify Chronic Liver Injury Biomarkers Following Exposure to Arsenic in Rats.

Biological trace element research·2022
Same author

Corrigendum to "SFPQ is involved in regulating arsenic-induced oxidative stress by interacting with the miRNA-induced silencing complexes" [Environ. Pollut. 261 (2020) 114160].

Environmental pollution (Barking, Essex : 1987)·2021
Same author

Computational Systems Pharmacology, Molecular Docking and Experiments Reveal the Protective Mechanism of Li-Da-Qian Mixture in the Treatment of Glomerulonephritis.

Journal of inflammation research·2021
Same author

Characteristics and phylogenetic analysis of the complete chloroplast genome of <i>Lilium concolor</i> Salisb. (Liliaceae) from Jilin, China.

Mitochondrial DNA. Part B, Resources·2021
This summary is machine-generated.

A new solid-phase catalysis method reduces sample loss and time for Nanopore direct RNA sequencing. This advance enables low-input RNA analysis for improved pathogen detection and transcript identification.

Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Nanopore direct RNA sequencing requires substantial RNA input (500 ng), limiting low-input applications like pathogen surveillance.
  • Current library preparation methods using solid-phase reversible immobilization (SPRI) beads cause significant sample loss, contributing to the high input requirement.

Purpose of the Study:

  • To develop a novel RNA library preparation strategy for Nanopore direct RNA sequencing that minimizes sample input requirements.
  • To overcome the limitations of SPRI bead purification in low-input RNA sequencing.

Main Methods:

  • A solid-phase catalysis approach was developed, immobilizing poly(A) polymerase and T4 DNA ligase for concurrent processing of non-polyadenylated transcripts.
  • The prepared library was directly loaded onto the Nanopore flow cell without SPRI bead purification.
Keywords:
Oxford Nanopore Technologiesdirect RNA-seqfoodborne pathogenimmobilized enzymesnext-generation sequencingtranscriptome

More Related Videos

High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR
11:00

High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

Published on: November 28, 2016

12.1K
Validating Whole Genome Nanopore Sequencing, using Usutu Virus as an Example
05:45

Validating Whole Genome Nanopore Sequencing, using Usutu Virus as an Example

Published on: March 11, 2020

9.0K

Related Experiment Videos

Last Updated: Oct 11, 2025

Sequencing of mRNA from Whole Blood using Nanopore Sequencing
11:26

Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Published on: June 3, 2019

14.0K
High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR
11:00

High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

Published on: November 28, 2016

12.1K
Validating Whole Genome Nanopore Sequencing, using Usutu Virus as an Example
05:45

Validating Whole Genome Nanopore Sequencing, using Usutu Virus as an Example

Published on: March 11, 2020

9.0K
  • Whole transcriptome sequencing was performed using Listeria monocytogenes as a model organism.

    • Main Results:

    • The novel method demonstrated minimal sample loss, reduced preparation time, and increased sequencing throughput compared to standard methods.
    • Reduced nanopore fouling was observed, leading to higher quality sequencing data.
    • The approach successfully enabled low-input Nanopore direct RNA sequencing, enhancing pathogen detection and transcript identification.

    Conclusions:

    • The solid-phase catalysis strategy significantly improves Nanopore direct RNA sequencing efficiency, particularly for low-input samples.
    • This method facilitates improved pathogen surveillance and microbial community analysis.
    • Accurate RNA consensus with high fidelity and identification of more expressed genes were achieved for both high and low RNA inputs.