Immunoglobulin Gene Amplification
Immunoglobulin Gene Amplification - Related Videos
02:48
Loop-Mediated Isothermal Amplification for Bacterial Gene Detection
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2025
Source: Domesle, K. J., et al. Loop-Mediated Isothermal Amplification for Screening Salmonella in Animal Food and Confirming Salmonella from Culture Isolation. J. Vis. Exp. (2020).This video demonstrates the loop-mediated isothermal amplification (LAMP) technique for detecting a target bacterial gene by amplifying DNA at a constant temperature. The reaction mixture is prepared by adding an isothermal master mix, specific primers for the target gene, and samples containing DNA templates. The...
09:44
Rapid Amplification of cDNA Ends
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2023
Source: Pablo Sanchez Bosch2, Sean Corcoran2 and Katja Brückner1,2,3 1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research 2Department of Cell and Tissue Biology, 3Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Rapid Amplification of cDNA Ends (RACE) is a technique that allows amplification of full-length cDNA from mRNA by extending to the 3’ or 5’ end, even without prior knowledge of the sequence (Frohman et al.,...
04:48
A Paper-Based Immunoassay for the Detection of Immunoglobulin G
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2025
The video demonstrates a paper-based immunoassay, involving amine-functionalized cellulose discs cross-linked with glutaraldehyde. Covalent immobilization of IgG-Fc-specific capture antibodies, followed by the introduction of immunoglobulin G and horseradish peroxidase-conjugated detection antibodies, results in a color change confirming the presence of immunoglobulin G.
03:31
A Microfluidic Cartridge-Based Detection System for Allergen-Specific Immunoglobulin E
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2025
This video demonstrates a protocol for detecting serum-specific immunoglobulin E using a microfluidic cartridge-based chemiluminescence system. Serum IgEs interact with distinct allergens immobilized in the reaction zone of the cartridge, which are subsequently detected with peroxidase-conjugated anti-IgE antibodies and a chemiluminescent substrate, allowing simultaneous detection of multiple allergen-specific IgEs.
02:41
Amplification of a Suicide Plasmid in a Bacterial Host
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2025
Source: Tomás, J. M., et al. Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility. J. Vis. Exp. (2022).This video demonstrates the initiator-dependent amplification of a suicide plasmid in E. coli. The plasmid, which carries an antibiotic resistance gene, replicates only in the presence of an initiator protein encoded by the host. After electroporation and incubation in recovery medium, the mixture is plated on antibiotic-containing agar.
FAQs
Primer design determines which immunoglobulin sequences are recovered. Primers bind conserved regions flanking variable genes, allowing amplification across the more diverse antibody-coding segment while retaining sequence information useful for comparison. This targeting focuses the reaction on immunoglobulin genes rather than unrelated genetic material, supporting downstream analysis of B-cell responses.
Repeated PCR cycles increase product abundance through three coordinated stages: denaturation separates template strands, primer annealing allows primers to bind their conserved targets, and extension copies the selected regions. Starting with limited genetic material, the process produces enough amplified sequence for sequencing or related downstream analysis.
Sequencing the amplified products can describe antibody repertoire diversity and identify clonotypes, meaning groups of related antibody sequences. It can also reveal variable regions that bind a particular antigen when those sequences are recovered for further use. These outputs connect molecular sequence data with patterns of B-cell immune responses.
A basic workflow starts with limited genetic material and uses primers directed at conserved regions surrounding immunoglobulin variable genes. PCR then generates many copies through repeated denaturation, annealing, and extension cycles. Researchers can sequence the resulting products to examine repertoire features, identify clonotypes, or recover variable regions for recombinant antibody production.
In immunology and infection research, amplified immunoglobulin sequences help track B-cell responses and examine pathogen-specific immunity. Comparing sequence patterns can support investigations of how antibody populations are represented in an immune response. The resulting information also contributes to diagnostic development and to studies aimed at creating therapeutic antibodies.
Amplification can recover antibody variable regions from limited genetic material, preserving sequence information associated with antigen binding. Those recovered regions provide a molecular starting point for recombinant antibody production. This application links analysis of naturally occurring B-cell responses with the development of antibody-based research, diagnostic, or therapeutic tools.