An Overview Of Types Of PCR Probes

A PCR probe is a DNA or RNA sequence with a reporter molecule that is very specific and recognizes sequences of the DNA or RNA. Probes are essential since they help observe a target gene. A probe determines whether a particular DNA is present in a sample since it interacts with the probe and the sample. DNA probes may be used to research certain diseases, such as cancer in a gene. Probes are also used to identify microorganisms present in an animal. A DNA molecule is a double strand held by nucleotide base pairs. Probes can also be used in the diagnosis of infections. DNA is mainly of the same composition and structures, making it hard to take an interest in a specific area of interest and making using a gene probe efficient. DNA is an excellent way of identifying organisms in a living thing, especially the small ones. There are different types of PCR probes, which have common characteristics. Characteristics like using fluorescence enable the probes to detect a target sequence by binding them.
This article will discuss the different types of probes and how they work.
Hydrolysis Probes
Hydrolysis probes consist of oligonucleotide, fluorescent reporter, and a quencher. They are QPCR probes that target specific sequences. In qPCR, the hydrolysis probe binds to the target sequence, where they are then separated from the reporter, which also separates it from the quencher. In the hydrolysis probe, the fluorescent and the quencher are placed at specific locations of the oligonucleotide, which connects the molecules, enabling them to perform reactions. These probes use hybridization, whereby the DNA bonds form a double-stranded molecule.
Dual Hybridization Probes
Dual hybridization probes consist of donor fluorophore, acceptor fluorophore, and double hybridization. In dual hybridization probes, there are two oligonucleotides, one carrying a donor fluorophore and the other having the acceptor. These fluorophores must contact the donor to release its energy for the acceptor to receive it. The oligonucleotides and the fluorophores link with the target sequence for the energy transfer, and then the acceptor releases fluorescence. This probe is excellent in the specificity of the target sequence.
Molecular Beacon Probes
A molecular beacon probe has a quencher, a reporter, and an oligonucleotide. In this type of probe, the quencher and fluorescent reporter are close together since the oligonucleotide is designed to form a stem and loop. Like hydrolysis probes, the probe binds to the target sequence, and then the quencher and reporter are separated, allowing the reporter to emit fluorescence. This probe can identify multiple target sequences in one go because of their specificity and multiplexing abilities.
Eclipse Probes
An eclipse probe has components such as an oligonucleotide, reporters, a quencher, and a miner groove binder. There is a distance between the quencher and the reporter, therefore favouring the fluorescence. The minor groove interacts with the DNA. This probe binds to the target sequence and then changes its structure, separating the quencher and the reporter, producing fluorescence.
In conclusion, using probes in medicine has helped in DNA sequencing, which helps diagnose and treat diseases. Targeting a DNA molecule also helps determine its varying and changes in mutation.