Conventional Pcr
Conventional Pcr - Related Videos
10:55
Detection of Tilapia Lake Virus Using Conventional RT-PCR and SYBR Green RT-qPCR
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Cited by 28 •
2018
This protocol diagnoses Tilapia Lake Virus (TiLV) in tilapia tissues using RT-PCR methodologies. The entire method is described from tissue dissection to total RNA extraction, followed by cDNA synthesis and detection of TiLV by either conventional PCR or quantitative PCR using dsDNA binding a fluorescent binding dye.
01:30
Sign Convention
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2025
When analyzing a beam subjected to various loads, it is crucial to understand the internal forces and moments generated within the structure. These internal forces can be broadly classified into normal forces, shear forces, and bending moments. To determine these forces and moments, we use the method of sections and apply a specific sign convention based on their direction and the side of the section being analyzed. The normal force acts perpendicular to the beam's cross-section and can cause...
08:51
Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR (RS-PCR)
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Cited by 9 •
2016
Here, ribosomal spacer PCR (RS-PCR) is used together with a miniaturized electrophoresis system as a fast and high resolution method for genotyping S. aureus at moderate costs allowing a high throughput.
01:15
Enolate Mechanism Conventions
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2023
When a carbonyl compound is treated with a strong base, the α position gets deprotonated to give a resonance-stabilized intermediate called an enolate. Enolates are ambident nucleophiles because they possess two nucleophilic sites that can attack an electrophile owing to the delocalization of the negative charge between the α carbon and oxygen atoms. When the oxygen atom attacks an electrophile, it is called O-attack, whereas electrophilic attack via the α carbon is known as C-attack. C-attack...
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.
FAQs
Primers determine which DNA region is copied by conventional PCR. During annealing, each primer binds a matching sequence, establishing the starting points for DNA synthesis and confining amplification to the selected target. This sequence-specific recognition is central to analyzing a chosen gene or detecting a pathogen-associated DNA sequence rather than copying all DNA in the sample.
Each cycle changes the DNA template into a form the polymerase can use. Denaturation separates the strands, annealing permits primers to bind, and extension uses nucleotides to build complementary strands. Repeating these stages causes the selected sequence to accumulate, so a target that was initially difficult to detect becomes available for downstream analysis.
Thermostable DNA polymerase is important because the reaction repeatedly passes through denaturation and extension conditions. The enzyme remains suitable for synthesis during this cycling process, while the primer-template match guides where complementary strands begin. Together, enzyme stability and sequence-specific priming allow the same target region to be copied through successive rounds rather than requiring a new synthesis setup for each cycle.
Conventional PCR produces an endpoint result after cycling is complete. Researchers commonly examine the amplified material with agarose gel electrophoresis, allowing them to assess whether the expected DNA product was generated. This readout supports applications such as checking cloning results, identifying target sequences, or detecting pathogen-related DNA after the amplification reaction.
A conventional PCR workflow starts with template DNA, sequence-specific primers, nucleotides, and a thermostable DNA polymerase. The reaction is placed in a thermal cycler, where denaturation, annealing, and extension are repeated. After cycling, researchers typically use agarose gel electrophoresis to examine the amplified material, linking the reaction itself to a visible analytical result.
Its applications include identifying genes, detecting pathogens, verifying cloning results, and analyzing genetic variation. The same amplification strategy can also prepare DNA for sequencing. Thus, researchers can adapt the technique to questions about the presence of a sequence, the success of a molecular biology procedure, or differences among DNA samples.
Within biology, the method links a defined DNA target to downstream genetic analysis. Amplified products can be examined after electrophoresis, used to verify cloning, analyzed for variation, or prepared for sequencing. Its value is therefore both analytical and preparative: it can provide evidence about a sequence and produce material for subsequent molecular investigations.