What is real-time PCR?

Quantitative PCR (qPCR), also called real-time PCR or quantitative real-time PCR, is a PCR-based technique that couples amplification of a target DNA sequence with quantification of the concentration of that DNA species in the reaction.

What is PCR and what is its function?

Polymerase chain reaction (PCR) is a laboratory technique used to amplify DNA sequences. The method involves using short DNA sequences called primers to select the portion of the genome to be amplified.

How is PCR technology used?

PCR is used in many research labs, and it also has practical applications in forensics, genetic testing, and diagnostics. For instance, PCR is used to amplify genes associated with genetic disorders from the DNA of patients (or from fetal DNA, in the case of prenatal testing).

What is real-time PCR describe the working principles of probe based real-time PCR assays?

Probe-based real-time PCR chemistry offers the benefit of low background fluorescence and high sensitivity to detect a single transcript copy. The principle of probe-based qPCR is based on the detection of complementary strand extension in real time, which involves hydrolysis of the fluorescently-tagged probe.

Why it is called real time PCR?

In order to robustly detect and quantify gene expression from small amounts of RNA, amplification of the gene transcript is necessary. This measurement is made after each amplification cycle, and this is the reason why this method is called real time PCR (that is, immediate or simultaneous PCR).

Why do we use PCR?

PCR is a common tool used in medical and biological research labs. It is used in the early stages of processing DNA for sequencing?, for detecting the presence or absence of a gene to help identify pathogens?during infection, and when generating forensic DNA profiles from tiny samples of DNA.

Why do we use real-time PCR?

The main advantage of real-time PCR over PCR is that real-time PCR allows you to determine the initial number of copies of template DNA (the amplification target sequence) with accuracy and high sensitivity over a wide dynamic range.

What is the function of the quencher in a DNA probe used in qPCR?

The quencher absorbs the light emission from the reporter when the probe is intact. During amplification, the probe binds to the target and the DNA Polymerase with a 5–>3 exonuclease activity cleaves the reporter dye from a probe emitting fluorescence.