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

11.0K
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...
11.0K
Pre-mRNA Processing: Modification of pre-mRNA Ends01:35

Pre-mRNA Processing: Modification of pre-mRNA Ends

12.9K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps...
12.9K
pre-mRNA Processing02:01

pre-mRNA Processing

55.4K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
55.4K
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

6.8K
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...
6.8K
Ribosome Profiling02:24

Ribosome Profiling

3.9K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.9K
Next-generation Sequencing03:00

Next-generation Sequencing

95.7K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
95.7K

You might also read

Related Articles

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

Sort by
Same author

A multi-omics approach to identify the impact of miR-411ed on NSCLC TKI resistance.

bioRxiv : the preprint server for biology·2026
Same author

A plasma miRNA-based classifier for small cell lung cancer diagnosis.

Frontiers in oncology·2023
Same author

A-to-I edited miR-411-5p targets MET and promotes TKI response in NSCLC-resistant cells.

Oncogene·2023
Same author

Cytoplasmic mRNA recapping has limited impact on proteome complexity.

Open biology·2020
Same author

Inhibition of cytoplasmic cap methylation identifies 5' TOP mRNAs as recapping targets and reveals recapping sites downstream of native 5' ends.

Nucleic acids research·2020
Same author

A recap of RNA recapping.

Wiley interdisciplinary reviews. RNA·2018
Same journal

Editorial: Technologies for RNA Detection.

Bio-protocol·2026
Same journal

One-Step Affinity Purification of MarathonRT Reverse Transcriptase for RNA Sequencing Applications.

Bio-protocol·2026
Same journal

Enhanced RNA-Seq Expression Profiling and Functional Enrichment in Non-model Organisms Using Custom Annotations.

Bio-protocol·2026
Same journal

Using Combined Fluorescent In Situ Hybridization With Immunohistochemistry to Co-localize mRNA in Diverse Neuronal Cell Types.

Bio-protocol·2026
Same journal

Stepwise Protocol for Alternative Splicing Analysis in Single-Cell SMART-Seq2 RNA-Seq Data.

Bio-protocol·2026
Same journal

Enriching Bacteria-Specific RNA From Host Samples Before NGS With Transcript-Capture.

Bio-protocol·2026
See all related articles

Related Experiment Video

Updated: Nov 15, 2025

3' End Sequencing Library Preparation with A-seq2
12:01

3' End Sequencing Library Preparation with A-seq2

Published on: October 10, 2017

10.9K

Analyzing (Re)Capping of mRNA Using Transcript Specific 5' End Sequencing.

Daniel Del Valle Morales1,2, Daniel R Schoenberg1,2

  • 1Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210, USA.

Bio-Protocol
|March 4, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a new method to locate 5' cap recapping events on eukaryotic messenger RNAs (mRNAs). The technique uses a specialized kit to tag and sequence these recapped mRNA ends, aiding functional studies.

Keywords:
5′ CapCap end mappingPCRPrimer ligationmRNA

More Related Videos

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

39.5K
Adapting 3' Rapid Amplification of CDNA Ends to Map Transcripts in Cancer
09:38

Adapting 3' Rapid Amplification of CDNA Ends to Map Transcripts in Cancer

Published on: March 28, 2018

12.6K

Related Experiment Videos

Last Updated: Nov 15, 2025

3' End Sequencing Library Preparation with A-seq2
12:01

3' End Sequencing Library Preparation with A-seq2

Published on: October 10, 2017

10.9K
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

39.5K
Adapting 3' Rapid Amplification of CDNA Ends to Map Transcripts in Cancer
09:38

Adapting 3' Rapid Amplification of CDNA Ends to Map Transcripts in Cancer

Published on: March 28, 2018

12.6K

Area of Science:

  • Molecular Biology
  • RNA Biology
  • Gene Expression

Background:

  • The 5' cap is essential for eukaryotic mRNA stability and translation.
  • Cytoplasmic recapping of mRNA occurs after decapping or cleavage, but its location and function are unclear.
  • Identifying recapping sites is crucial for understanding mRNA processing and regulation.

Purpose of the Study:

  • To develop and validate a method for identifying and locating 5' cap recapping events on eukaryotic mRNAs.
  • To provide a tool for investigating the functional consequences of cytoplasmic mRNA recapping.

Main Methods:

  • Utilized the Lexogen TeloPrime cDNA synthesis kit for specific tagging of recapped 5' ends.
  • Employed a proprietary DNA ligase to append a DNA oligonucleotide to cDNA hybridized to mRNA.
  • Used PCR amplification and direct sequencing (Sanger or Illumina) to determine the junction site between the tag and the mRNA.

Main Results:

  • Successfully developed an assay to tag and sequence recapped 5' ends of eukaryotic mRNAs.
  • The method precisely identifies the location of recapping events on target transcripts.
  • Demonstrated applicability to all capped transcripts, adaptable for various sequencing scales.

Conclusions:

  • The presented approach enables accurate mapping of cytoplasmic mRNA recapping events.
  • This technique facilitates research into the functional significance of mRNA recapping.
  • Offers a versatile tool for studying mRNA 5' end processing in eukaryotes.