Thermocycling
Thermocycling - Related Videos
09:02
Rapid PCR Thermocycling using Microscale Thermal Convection
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Cited by 21 •
2011
We describe a novel method to perform DNA replication via the polymerase chain reaction (PCR). Thermal convection is harnessed to continuously shuttle reagents between denaturing, annealing, and extension conditions by maintaining opposing surfaces of the reactor at constant temperature. This inherently simple design promises to make rapid PCR more accessible.
04:23
Standardized Protocol to Evaluate the Effect of Surface Treatments, Luting Cements, and Thermocycling on PEEK–Composite Bond Strength
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2026
This protocol aims to evaluate the effects of surface treatment protocols, luting cement selection, and thermocycling on the shear bond strength between PEEK and an indirect composite resin.
06:55
Simple Bulk Readout of Digital Nucleic Acid Quantification Assays
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Cited by 5 •
2015
We describe an endpoint digital assay for quantifying nucleic acids with a simplified (analog) readout. We measure bulk fluorescence of droplet-based digital assays using a standard qPCR machine rather than specialized instrumentation and confirm our results by microscopy.
10:37
Perturbations of Circulating miRNAs in Irritable Bowel Syndrome Detected Using a Multiplexed High-throughput Gene Expression Platform
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Cited by 6 •
2016
We describe the use of a multiplexed high-throughput gene expression platform that quantitates gene expression by barcoding and counting molecules in biological substrates without the need for amplification. We used the platform to quantitate microRNA (miRNA) expression in whole blood in subjects with and without irritable bowel syndrome.
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...
FAQs
Each PCR stage prepares the reaction for the next one. Denaturation makes DNA strands accessible, annealing positions primers on complementary sequences, and extension lets the heat-stable polymerase copy from those primers. Because newly synthesized strands become templates in later cycles, repeating the sequence increases the amount of the selected region, enabling analysis from limited starting DNA.
The heat-stable polymerase remains functional through the repeated high-temperature conditions used to separate DNA strands. A polymerase that lost activity during these cycles could not continue synthesizing DNA during extension. Its persistence allows the same reaction mixture to support repeated copying, which is essential for producing amplified material for downstream biological analyses.
Primer binding provides sequence-level control over amplification. During annealing, each primer binds a complementary DNA sequence and establishes a starting point for polymerase-mediated synthesis. The arrangement of the primer-target sequences identifies the region copied during extension, supporting targeted applications such as genotyping, pathogen detection, cloning, and DNA sequencing workflows.
A researcher places the PCR reaction in a thermocycler and programs repeated temperature changes for denaturation, annealing, and extension. The cycle is repeated to generate more copies of the target region. After amplification, the resulting DNA can support genotyping, cloning, sequencing workflows, pathogen detection, or molecular diagnostic analysis.
Thermocycling is particularly valuable when the available genetic material is small or limited. PCR amplification increases the amount of a selected DNA region before interpretation, allowing analysis from samples that provide little starting template. This capability supports sensitive biological applications, including genotyping, pathogen detection, molecular diagnostics, cloning, and sequencing workflows.
Amplified DNA from this process can support several distinct biological workflows. Researchers may use it for genotyping, detecting pathogens, preparing DNA for cloning, or contributing material to sequencing workflows. Molecular diagnostics also rely on the ability to analyze selected genetic regions, making thermocycling a flexible step across multiple forms of biological investigation.