10 Nisan 2011 Pazar

Real-time PCR

In the past, the PCR was carried out of the tube and the reaction is complete when the products of the reaction (the amplified DNA fragments) is analyzed by gel electrophoresis and visualized. However, the Real-Time PCR enables the analysis of the products, while the reaction is really going on. This is achieved by different fluorescent dyes that react to the amplified product can be measured with an instrument. This facilitates a quantitative DNA. Not just immediately say "our" DNA sample, but the "how". Quantitative PCR (Q-PCR) technique, as is known, the amount used in the PCR product (usually a real-time PCR process) measurement. This is the method of choice for quantitative measurement of the starting amount of DNA, cDNA or RNA. PCR is often used to determine whether the DNA sequence present in the sample, and the number of copies of the sample. Another advantage of the Real-Time PCR test for speed, because it is not necessary for electrophoresis, or other proceeding shall be done after the DNA amplification reaction.

The development methods of the sealed tube fluorescent polymerase chain reaction greatly simplified the process of quantification. The current approach is to use fluorescent probes that interact with the PCR amplification product of the kinetic measurements of accumulation possible. These methods are a general approach to quantitative DNA probe as a DNA-binding fluorescent dyes. There are a few carefully-specific probes, that the phenomenon of fluorescence resonance energy transfer (FRET) to use. Development of tools to deliver real-time monitoring of fluorescence within the PCR reaction allowed the ship is a very important step in the PCR. The technology is very flexible and many alternative instruments and fluorescent probe systems currently available. Real-time PCR can be completed very quickly, since no manipulations are required after amplification. Identification of the amplification products of the probe detection in real-time analysis of very precise size of the gels. Analysis of the progress of the reaction sequence allows the exact quantification of a wide dynamic range, provided that appropriate standards are available. Further investigation of the real-time PCR products of the initial reaction mixture, using probes and melting analysis can detect sequence variants including single base mutations. Real-time PCR applications in many areas of biological science. Applications include the analysis of gene expression, the diagnosis of infectious diseases and human genetic studies. As a result, the devices in real-time fluorimetry machines are also compatible with alternative methods such as the reinforcement provided lishing, fluorescent endpoint is available.

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