Polymerase Amplification
Polymerase Amplification - Related Videos
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.
13:27
PCR: The Polymerase Chain Reaction
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2023
The polymerase chain reaction, or PCR, is a technique used to amplify DNA through thermocycling – cyles of temperature changes at fixed time intervals. Using a thermostable DNA polymerase, PCR can create numerous copies of DNA from DNA building blocks called dNTPs. There are three steps in PCR: denaturation, annealing, and elongation. Denaturation is the first step in the cycle and causes the DNA to melt by disrupting hydrogen bonds between the bases resulting in single-stranded DNA. Annealing...
06:47
Combined Recombinase Polymerase Amplification CRISPR/Cas12a Assay for Detecting Fusarium oxysporum f. sp. cubense Tropical Race 4
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2025
In this protocol, a combined recombinase polymerase amplification CRISPR/Cas12a assay is illustrated for the detection of the invasive plant pathogen, Fusarium oxysporum f. sp. cubense tropical race 4.
02:10
Translesion DNA Polymerases
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2020
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication. TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
00:58
Eukaryotic RNA Polymerases
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2025
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action. All three eukaryotic RNAPs require specific transcription factors, of which the...
FAQs
Sequence-specific primers identify the DNA region to be copied by binding to complementary sequences. The thermostable DNA polymerase then extends from those primers to build new complementary strands. Together, these components provide both targeting and strand synthesis, allowing researchers to focus amplification on a selected sequence rather than copying genetic material indiscriminately.
The temperature cycles coordinate three essential events: heating separates the two DNA strands, cooling permits sequence-specific primers to bind, and polymerase extension creates complementary strands. Repeating this sequence increases the amount of the selected DNA region. Controlled cycling therefore connects strand separation, recognition, and synthesis into one amplification process.
The technique can support analysis of genetic variants and DNA sequences while also measuring nucleic acids associated with gene expression. Examining these molecular signals provides complementary information about inherited or acquired sequence differences and regulation-related activity. In neuroscience, that combination helps researchers investigate how molecular changes relate to neural development, function, and disease.
A basic workflow requires genetic material containing the target sequence, sequence-specific primers, a thermostable DNA polymerase, and controlled heating and cooling. The process begins by separating DNA strands, followed by primer binding and polymerase extension. Repeating these stages generates enough target material for detection, analysis, or comparison with other samples.
Researchers can apply the method when neural samples contain only very small amounts of genetic material or when a specific sequence must be examined. Supported uses include genotyping, analyzing neural tissue, investigating genetic variants, and measuring nucleic acids associated with gene expression. These applications make molecular signals accessible for studies of neural biology and disease.
Polymerase amplification can make selected genetic material detectable and suitable for analysis, including DNA sequences from neural tissue and signals associated with gene expression. Researchers can use these results to identify or compare genetic variants, examine molecular patterns, and validate other molecular findings. The resulting evidence helps relate sequence and regulation to neural development, function, and disease.