Fluorescent Primers
Fluorescent Primers - Related Videos
11:14
Primer-Free Aptamer Selection Using A Random DNA Library
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Cited by 20 •
2010
SELEX protocols comprise multiple rounds of selection, each of which require regeneration of bound ligands, which in turn require fixed primer sequences flanking the random library regions. These fixed primer sequences can interfere with the selection process (false positives and negatives). Here we present a primer-free protocol.
10:51
Fluorescence Based Primer Extension Technique to Determine Transcriptional Starting Points and Cleavage Sites of RNases In Vivo
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Cited by 17 •
2014
We here describe a fluorescence based primer extension method to determine transcriptional starting points from bacterial transcripts and RNA processing in vivo using an automated gel sequencer.
15:28
Primer Extension Capture: Targeted Sequence Retrieval from Heavily Degraded DNA Sources
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Cited by 41 •
2009
We present a method of targeted ancient DNA sequence retrieval, which we used to reconstruct the complete mitochondrial genomes of five Neandertal individuals. Comparison of these sequences with present day humans suggests that Neandertals had a long term low effective population size.
10:17
Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers
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Cited by 7 •
2017
Most common gut content analyses of macroinvertebrates are visual. Requiring intense knowledge about morphological diversity of prey organisms, they miss soft bodied prey and, due to strong comminution of prey, are nearly impossible for some organisms, including amphipods. We provide detailed, novel genetic approaches for macroinvertebrate prey identification in the diet of amphipods.
07:30
Primer for Immunohistochemistry on Cryosectioned Rat Brain Tissue: Example Staining for Microglia and Neurons
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Cited by 28 •
2015
This introductory level protocol describes the reagents, equipment, and techniques required to complete immunohistochemical staining of rodent brains, using markers for microglia and neuronal elements as an example.
FAQs
A fluorescent primer contributes both target recognition and signal generation. Its sequence anneals to a complementary region, allowing DNA polymerase to extend the primer and produce labeled amplicons. When the fluorophore is excited, the resulting light provides a readout linked to the amplified genetic target, making detection more direct than relying only on an unlabeled endpoint product.
Sequence-specific annealing determines which genetic region receives the fluorescent label during amplification. A primer that matches the intended template supports extension of that target, while the resulting signal reflects the selected sequence rather than an unspecified DNA product. This targeting is especially relevant for distinguishing pathogen sequences, genotypes, or immune-receptor sequences in complex samples.
The same fluorescently labeled amplification strategy can support different readout formats. Capillary electrophoresis resolves amplified products so their associated fluorescent signals can be detected, whereas fluorescence-based assays monitor the signal during amplification. The choice therefore affects when and how the labeled products are observed, while both approaches use light to assess target-specific amplification.
Fluorescence supplies a measurable optical signal from the labeled amplification products rather than requiring interpretation based solely on a final product. This supports sensitive, target-specific measurements and can make amplified genetic targets easier to detect. In infection and immunology studies, that added measurement capability helps connect a detected sequence with pathogen or immune-related analysis.
The workflow begins with primer annealing to the complementary template, followed by DNA polymerase extension and production of fluorescently labeled amplicons. The fluorophore is then excited, and the emitted signal is detected either through capillary electrophoresis or a fluorescence-based assay. This sequence links target selection, amplification, labeling, and optical measurement in one analysis.
For infectious-disease studies, primers directed at selected pathogen sequences can label amplified targets for sensitive detection. The resulting fluorescence can also support genotyping by distinguishing measurements associated with different genetic targets. These uses provide sequence-focused information that helps researchers identify infectious agents and examine genetic variation relevant to infection research.
In immunology, fluorescent primers can label amplified immune-receptor sequences for repertoire analysis, allowing researchers to measure target-specific genetic patterns associated with host responses. They also support multiplex testing, in which more than one target can be examined within an assay context. These applications connect fluorescent detection with broader studies of immune response and infectious disease.