Amplification Reaction System
Amplification Reaction System - Related Videos
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.,...
10:12
Protein Misfolding Cyclic Amplification of Prions
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Cited by 7 •
2012
Protein misfolding cyclic amplification (PMCA) is an in vitro assay for the study of prion conversion and strain and species barriers. It can also be used as a prion detection assay.
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...
07:35
Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray
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Cited by 4 •
2014
An amplification microarray combines asymmetric PCR amplification and microarray hybridization into a single chamber, which significantly streamlines microarray workflow for the end user. Simplifying microarray workflow is a necessary first step for creating microarray-based diagnostics that can be routinely used in lower-resource environments.
08:37
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
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Cited by 17 •
2015
Provided is a protocol for developing a real-time recombinase polymerase amplification assay to quantify initial concentration of DNA samples using either a thermal cycler or a microscope and stage heater. Also described is the development of an internal positive control. Scripts are provided for processing raw real-time fluorescence data.
FAQs
Each cycle separates existing nucleic-acid strands, allows sequence-specific primers to attach, and lets the polymerase extend new strands using supplied nucleotides. The newly produced copies become templates in later cycles, so target enrichment increases exponentially rather than by a fixed amount. This amplification makes genetic material measurable even when the starting sample contains only a small quantity.
Primers determine which nucleic-acid sequence can be copied because they anneal to matching target regions before polymerase extension. Their sequence-specific binding helps distinguish the selected genetic material from unrelated sequences in the sample. Consequently, primer selection is central to detecting a particular pathogen genome or immune-related marker rather than amplifying nucleic acid indiscriminately.
RNA targets first require conversion into complementary DNA so they can enter the nucleic-acid amplification workflow described for DNA templates. This added step changes the form of the target without changing the goal of sequence enrichment. It allows researchers to examine RNA-associated pathogen or immune markers using subsequent primer-directed copying and repeated amplification cycles.
The system depends on carefully controlled conditions that coordinate strand separation, primer annealing, and polymerase-driven extension. If these stages do not occur in the intended sequence or under suitable conditions, target copying may be reduced or less selective. Proper control therefore supports the two key outcomes emphasized for these assays: sensitivity for scarce material and specificity for the selected sequence.
A basic workflow combines the sample template with sequence-specific primers, nucleotides, polymerase, and the required controlled reaction conditions. For an RNA target, complementary-DNA conversion precedes the amplification cycles. The system then repeatedly separates strands, permits primer annealing, and extends the primers enzymatically, producing enriched target material that can be measured or interpreted.
They are useful when investigators need to detect pathogen genomes, estimate microbial burden, or examine genetic material in clinical or experimental samples. Because amplification can make scarce targets measurable, the approach supports early detection and outbreak investigation. It also provides a molecular route for studying infection-related changes alongside other observations of host-pathogen responses.
In immunology and infection research, the same assay framework can identify immune-related nucleic-acid markers as well as pathogen genetic material. Researchers can therefore compare evidence of the infectious agent with molecular indicators of the host response. These measurements help evaluate host-pathogen interactions and support investigation of how infection and immunity are reflected in experimental or clinical samples.