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

10.4K
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...
10.4K
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

6.5K
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...
6.5K

You might also read

Related Articles

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

Sort by
Same author

Accelerated apolipoprotein A-II senile amyloidosis in a plasminogen activator inhibitor-1 knock-out model.

Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis·2026
Same author

Recurrence of light chain crystalline podocytopathy and tubulopathy after kidney transplantation achieving remission with an antimyeloma regimen.

American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons·2025
Same author

Untwisted amyloid fibrils from a transgenic mouse model of AL amyloidosis.

Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis·2025
Same author

DIS3 licenses B cells for plasma cell differentiation in humans.

Cellular & molecular immunology·2025
Same author

Revisiting proliferative glomerulonephritis with monoclonal immunoglobulin deposits through immunoglobulin repertoire sequencing.

Kidney international·2025
Same author

Guidelines for RNA Analysis by Reverse Transcription Quantitative Polymerase Chain Reaction.

Methods in molecular biology (Clifton, N.J.)·2025

Related Experiment Video

Updated: Sep 14, 2025

Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing
08:51

Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing

Published on: March 15, 2019

12.5K

5'RACE Method for RNA-Based Sequencing of the Mouse Immunoglobulin Repertoire.

Sandy Al Hayek1, Antoine Praité1, Jean-Marie Lambert1,2

  • 1UMR CNRS 7276, Inserm 1262, Université de Limoges, Limoges, France; CRIBL lab (Control of the B-cell Immune Response and Lymphoproliferations), Limoges, France; Team 3, BioPIC (Biology of Plasma Cells, Immunopathology and Cancer), Limoges, France.

Methods in Molecular Biology (Clifton, N.J.)
|July 23, 2025
PubMed
Summary

This study details a method for high-throughput sequencing of mouse immunoglobulin repertoires. The technique analyzes adaptive immune responses by examining B cell diversity and somatic hypermutations.

Keywords:
5′RACEImmunoglobulin repertoireNext-generation sequencingPaired-end sequencingRNAUnique molecular identifiers (UMI)

More Related Videos

Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell Differentiation
11:06

Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell Differentiation

Published on: September 20, 2017

6.2K
Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis
10:31

Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis

Published on: May 10, 2019

20.1K

Related Experiment Videos

Last Updated: Sep 14, 2025

Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing
08:51

Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing

Published on: March 15, 2019

12.5K
Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell Differentiation
11:06

Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell Differentiation

Published on: September 20, 2017

6.2K
Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis
10:31

Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis

Published on: May 10, 2019

20.1K

Area of Science:

  • Immunology
  • Molecular Biology
  • Genomics

Background:

  • The adaptive immune system relies on immunoglobulin (Ig) diversity for pathogen recognition.
  • Characterizing the Ig repertoire is crucial for understanding immune responses and developing targeted therapies.
  • Existing methods may lack the throughput or accuracy needed for comprehensive repertoire analysis.

Purpose of the Study:

  • To present a detailed methodology for high-throughput sequencing of the mouse Ig repertoire.
  • To enable comprehensive analysis of Ig diversity, clonality, and somatic hypermutations.
  • To provide insights into the adaptive immune response in various immunological contexts.

Main Methods:

  • RNA extraction from splenic B cells.
  • cDNA synthesis using 5'RACE (Rapid Amplification of cDNA Ends) and PCR amplification of Ig heavy and light chains.
  • Library preparation for Illumina next-generation sequencing (NGS) incorporating unique molecular identifiers (UMIs).

Main Results:

  • Successful amplification of Ighm, Ighg subclasses, and Igκ light chain.
  • Implementation of stringent quality control for RNA integrity and amplification accuracy.
  • Demonstration of the method's utility in analyzing Ig repertoire diversity and clonality.

Conclusions:

  • RNA-based 5'RACE-RepSeq is a powerful tool for high-throughput characterization of the mouse Ig repertoire.
  • This method provides valuable insights into somatic hypermutations and the adaptive immune response.
  • The protocol facilitates detailed analysis of Ig repertoire in diverse immunological research settings.