Realtime PCR for quantitative analysis from "summary" of Molecular Cloning by Joseph Sambrook,David William Russell
Real-time PCR is a powerful technique for quantitative analysis of nucleic acids. It is based on the polymerase chain reaction, in which a specific DNA sequence is amplified exponentially. However, in real-time PCR, the amplification of DNA is monitored in "real time" as the reaction is taking place, allowing for the quantification of the starting amount of DNA. One of the key components of real-time PCR is the use of fluorescent dyes or probes that bind specifically to the amplified DNA. These dyes emit a fluorescent signal that can be detected by a thermal cycler in each cycle of the PCR reaction. The amount of fluorescence is directly proportional to the amount of DNA being amplified, allowing for the quantification of the initial DNA concentration. There are several different methods for detecting the fluorescence in real-time PCR, including SYBR Green dye, TaqMan probes, and molecular beacons. Each of these methods has its own advantages and limitations, but all of them allow for highly sensitive and accurate quantification of nucleic acids. Real-time PCR can be used for a wide range of applications, including gene expression analysis, viral load quantification, and detection of genetic mutations. It is a versatile and reliable technique that has revolutionized the field of molecular biology.- Real-time PCR for quantitative analysis is a crucial tool for researchers studying nucleic acids. Its ability to provide accurate and sensitive quantification of DNA makes it an indispensable technique in many areas of molecular biology.
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