UPPCR stands for Universal Probe-based Polymerase Chain Reaction. It is a specific molecular biology technique that combines two well-known laboratory methods: the polymerase chain reaction (PCR) and the use of probes.
PCR is a technique used to amplify a specific segment of DNA, creating multiple copies of the target DNA sequence. It involves a series of temperature cycles, which allow for the denaturation, annealing, and extension of DNA sequences using a special DNA polymerase enzyme.
UPPCR enhances PCR by incorporating the use of probes. Probes are short, single-stranded DNA or RNA molecules that are complementary to the target DNA sequence. They are chemically modified to contain a fluorescent dye and a quencher molecule. The probe binds specifically to the target DNA sequence, providing a more accurate and specific detection method.
During UPPCR, the target DNA is first denatured into single strands and then mixed with PCR primers and probes. The primers initiate DNA replication, while the probes bind to the complementary DNA sequences. As the DNA amplification occurs, the probe is cleaved, separating the fluorescent dye from the quencher molecule. This allows for the detection and quantification of the amplified target DNA.
UPPCR is widely used in research and diagnostic laboratories for various applications, such as genetic testing, viral load quantification, and detecting specific mutations or gene expression levels. Its high specificity, sensitivity, and ability to quantify DNA make it an essential tool in many areas of molecular biology.
