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 Concept Videos

RNA-seq03:21

RNA-seq

10.0K
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...
10.0K

You might also read

Related Articles

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

Sort by
Same author

Single-cell exon deletion profiling reveals splicing events that shape gene expression and cell state dynamics.

Nature communications·2026
Same author

RNA-coupled CRISPR screens reveal ZNF207 as a regulator of LMNA aberrant splicing in progeria.

Molecular cell·2025
Same author

RNA-coupled CRISPR Screens Reveal ZNF207 as a Regulator of LMNA Aberrant Splicing in Progeria.

bioRxiv : the preprint server for biology·2025
Same author

A ligation-independent sequencing method reveals tRNA-derived RNAs with blocked 3' termini.

Molecular cell·2024
Same author

<i>CircHTT(2,3,4,5,6)</i> <i>-</i> co-evolving with the <i>HTT</i> CAG-repeat tract - modulates Huntington's disease phenotypes.

Molecular therapy. Nucleic acids·2024
Same author

Genome-scale exon perturbation screens uncover exons critical for cell fitness.

Molecular cell·2024
Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Jul 2, 2025

Identification of Circular RNAs using RNA Sequencing
08:25

Identification of Circular RNAs using RNA Sequencing

Published on: November 14, 2019

12.2K

Purification of Circular RNAs Using Poly(A) Tailing Followed by RNase R Digestion.

Mei-Sheng Xiao1, Jeremy E Wilusz2

  • 1RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 21, 2024
PubMed
Summary
This summary is machine-generated.

This study introduces an improved method for purifying circular RNAs (circRNAs) from total RNA. The enhanced protocol increases circRNA enrichment, aiding in the identification and validation of circRNAs for functional studies.

Keywords:
BacksplicingCIRCexplorer2CIRI2CiRNACircRNAG-quadruplexPoly(A) polymeraseRNA-seqRT-qPCRRibonuclease R

More Related Videos

Quantification of Circular RNAs Using Digital Droplet PCR
08:39

Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

3.5K
Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA
14:49

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Published on: October 27, 2011

39.2K

Related Experiment Videos

Last Updated: Jul 2, 2025

Identification of Circular RNAs using RNA Sequencing
08:25

Identification of Circular RNAs using RNA Sequencing

Published on: November 14, 2019

12.2K
Quantification of Circular RNAs Using Digital Droplet PCR
08:39

Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

3.5K
Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA
14:49

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA

Published on: October 27, 2011

39.2K

Area of Science:

  • Molecular Biology
  • Genomics
  • RNA Biology

Background:

  • Alternative splicing generates both linear mRNAs and circular RNAs (circRNAs) in eukaryotes.
  • Most circRNAs are expressed at low levels, making their detection challenging in standard RNA sequencing.
  • Ribonuclease R (RNase R) is commonly used to enrich circRNAs but struggles with linear RNAs possessing structured 3' ends or G-quadruplexes.

Purpose of the Study:

  • To develop an improved protocol for circRNA purification from total RNA.
  • To enhance the enrichment efficiency of circRNAs compared to standard RNase R digestion.
  • To facilitate the identification and validation of circRNAs for subsequent functional studies.

Main Methods:

  • An improved circRNA purification protocol involving a poly(A) tailing step before RNase R digestion.
  • RNase R digestion performed in a Li+ containing buffer to destabilize G-quadruplexes.
  • Quantitative RT-PCR (RT-qPCR) and RNA sequencing library generation for circRNA analysis.
  • Step-by-step methods for circRNA annotation using CIRI2 and CIRCexplorer2 algorithms.

Main Results:

  • The improved protocol achieves a two- to threefold higher enrichment of circRNAs compared to standard RNase R methods.
  • More efficient removal of linear RNAs, including those with structured 3' ends or G-quadruplexes.
  • Enables robust examination of individual circRNA expression and definition of the entire expressed circRNA set.

Conclusions:

  • The described biochemical method significantly improves circRNA purification efficiency.
  • This approach allows for the enrichment of circRNAs from any total RNA sample.
  • Facilitates rapid identification and validation of circRNAs for functional investigations.