RTqPCRmenetelmässä

RTqPCRmenetelmässä, often referred to as reverse transcription quantitative polymerase chain reaction, is a laboratory technique used to detect and quantify RNA levels. It combines the principles of reverse transcription and quantitative PCR. The process begins with RNA, which is first converted into complementary DNA (cDNA) using an enzyme called reverse transcriptase. This cDNA then serves as the template for quantitative PCR. In qPCR, a DNA polymerase amplifies specific DNA sequences. The "quantitative" aspect comes from the use of fluorescent dyes or probes that bind to the amplified DNA, allowing for real-time monitoring of the amplification process. As the PCR progresses, the fluorescence signal increases proportionally to the amount of amplified DNA. By comparing the fluorescence intensity to a standard curve or known control samples, researchers can determine the initial amount of RNA present in the original sample. This method is widely used in molecular biology research, diagnostics, and drug development for applications such as gene expression analysis, viral load quantification, and pathogen detection. Its sensitivity and specificity make it a powerful tool for studying biological processes at the molecular level.