Various detection and capture steps are involved in each approach to assess the amplification product, or amplicon. Generally, these techniques incorporate a capture step and a detection step, although how these steps are incorporated depends on the individual assay.Įxamples include the Amplicor HIV-1 Monitor Assay ( RT-PCR), which has the capacity to recognize HIV in plasma the HIV-1 QT ( NASBA), which is used to measure plasma viral load by amplifying a segment of the HIV RNA and transcription mediated amplification, which employs a hybridization protection assay to distinguish Chlamydia trachomatis infections. Irrespective of the approach used to amplify the amplicons, some technique must be used to quantitate the amplified product. This works similarly in the fungal domain with the 18S rRNA gene as well as the ITS1 non-coding region. For example, using the 16S rRNA gene, which is part of every bacterial and archaeal genome and is highly conserved, bacteria can be taxonomically classified by comparison of the amplicon sequence to known sequences. ĭNA sequencing technologies such as next-generation sequencing have made it possible to study amplicons in genome biology and genetics, including cancer genetics research, phylogenetic research, and human genetics. Technology Īnalysis of amplicons has been made possible by the development of amplification methods such as PCR, and increasingly by cheaper and more high-throughput technologies for DNA sequencing or next-generation sequencing, such as ion semiconductor sequencing, popularly referred to as the brand of the developer, Ion Torrent. ĭuring artificial amplification, amplicon length is dictated by the experimental goals. Circular amplicons consist of imperfect inverted duplications annealed into a circle and are thought to arise from precursor linear amplicons. Structure Īmplicons in general are direct repeat (head-to-tail) or inverted repeat (head-to-head or tail-to-tail) genetic sequences, and can be either linear or circular in structure. Amplification of the genes encoded by these amplicons generally increases transcription of those genes and ultimately the volume of associated proteins. In this context the term amplicon can refer both to a section of chromosomal DNA that has been excised, amplified, and reinserted elsewhere in the genome, and to a fragment of extrachromosomal DNA known as a double minute, each of which can be composed of one or more genes. It is also implicated in several forms of human cancer including primary mediastinal B cell lymphoma and Hodgkin's lymphoma. Natural gene duplication plays a major role in evolution. As it refers to the product of an amplification reaction, amplicon is used interchangeably with common laboratory terms, such as "PCR product."Īrtificial amplification is used in research, forensics, and medicine for purposes that include detection and quantification of infectious agents, identification of human remains, and extracting genotypes from human hair. In this context, amplification refers to the production of one or more copies of a genetic fragment or target sequence, specifically the amplicon. It can be formed artificially, using various methods including polymerase chain reactions (PCR) or ligase chain reactions (LCR), or naturally through gene duplication. In molecular biology, an amplicon is a piece of DNA or RNA that is the source and/or product of amplification or replication events.
The target sequence to be amplified is colored green. An amplicon sequence template that has been prepared for amplification.
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