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A strip of eight PCR tubes, each containing a 100 μL reaction mixture Placing a strip of eight PCR tubes into a thermal cycler. The polymerase chain reaction (PCR) is a method widely used to make millions to billions of copies of a specific DNA sample rapidly, allowing scientists to amplify a very small sample of DNA (or a part of it) sufficiently to enable detailed study.
The PCR method is extremely sensitive, requiring only a few DNA molecules in a single reaction for amplification across several orders of magnitude. Therefore, adequate measures to avoid contamination from any DNA present in the lab environment (bacteria, viruses, or human sources) are required.
Multiplex-PCR consists of multiple primer sets within a single PCR mixture to produce amplicons of varying sizes that are specific to different DNA sequences. By targeting multiple sequences at once, additional information may be gained from a single test run that otherwise would require several times the reagents and more time to perform.
Random amplified polymorphic DNA (RAPD), pronounced "rapid", [1] is a type of polymerase chain reaction (PCR), but the segments of DNA that are amplified are random. [2] The scientist performing RAPD creates several arbitrary, short primers (10–12 nucleotides), then proceeds with the PCR using a large template of genomic DNA, hoping that fragments will amplify.
Several DNA polymerases have been described with distinct properties that define their specific utilisation in a PCR, in real-time PCR or in an isothermal amplification. Being DNA polymerases, the thermostable DNA polymerases all have a 5'→3' polymerase activity, and either a 5'→3' or a 3'→5' exonuclease activity.
In silico PCR example result with FastPCR [7] [8] software. The design of appropriate short or long primer pairs is only one goal of PCR product prediction. Other information provided by in silico PCR tools may include determining primer location, orientation, length of each amplicon , simulation of electrophoretic mobility, identification of ...
In research or diagnosis DNA amplification can be conducted through methods such as: Polymerase chain reaction, an easy, cheap, and reliable way to repeatedly replicate a focused segment of DNA by polymerizing nucleotides, a concept which is applicable to numerous fields in modern biology and related sciences. [2]
A real-time polymerase chain reaction (real-time PCR, or qPCR when used quantitatively) is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR (i.e., in real time), not at its end, as in conventional PCR. Real-time PCR can be used ...
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