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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.
Chimeric polymerases overcome many limitations of native enzymes and are used in direct PCR amplification from cell cultures and even food samples, thus by-passing laborious DNA isolation steps. A robust strand-displacement activity of the hybrid TopoTaq polymerase helps solve PCR problems that can be caused by hairpins and G-loaded double ...
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 ...
PCR inhibitors are any factor which prevent the amplification of nucleic acids through the polymerase chain reaction (PCR). [1] PCR inhibition is the most common cause of amplification failure when sufficient copies of DNA are present. [2] PCR inhibitors usually affect PCR through interaction with DNA or interference with the DNA polymerase.
Later, the PCR process was adapted to the use of thermostable DNA polymerase from Thermus aquaticus, which greatly simplified the design of the thermal cycler. While in some old machines the block is submerged in an oil bath to control temperature, in modern PCR machines a Peltier element is commonly used.
RT-PCR is commonly used in research methods to measure gene expression. For example, Lin et al. used qRT-PCR to measure expression of Gal genes in yeast cells. First, Lin et al. engineered a mutation of a protein suspected to participate in the regulation of Gal genes. This mutation was hypothesized to selectively abolish Gal expression.
A quantitative PCR instrument [1] is a machine that amplifies and detects DNA. It combines the functions of a thermal cycler and a fluorimeter , enabling the process of quantitative PCR . The first quantitative PCR machine was described in 1993, [ 2 ] and two commercial models became available in 1996.
DNA spiking can also refer to a spike control in PCR, which is when DNA is added to a sample that will provide some signal (e.g. a plasmid or some synthetic DNA with a specific known sequence) to a reaction, and seeing if the reaction will amplify. This method is used to discover if the PCR method is working correctly, as in a PCR machine it ...
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