Do you know what is the RT-PCR Full Form? RT-PCR Full Form is Real-Time Reverse Transcription-Polymerase Chain Reaction. A scientist can decode the information of RNA, and viruses through the Reverse Transcription-Polymerase Chain Reaction technique.
One method for finding viral RNA in the human body is the reverse transcription-polymerase chain reaction. It is also employed to research the genetic composition of viruses. The human herpes virus, for example, may be present in that many persons.
The RT-PCR method was initially introduced in 1983 by Kary Mullis. The RT-PCR technique replaced the Northern Blot technique. Despite its drawbacks, the northern blot method, developed in 1977, was frequently employed to measure RNA. For purposes of detection, it should contain a significant amount of RNA.
What is the RT-PCR Full Form?
RT-PCR Full Form: Real-Time Reverse Transcription-Polymerase Chain Reaction
What is the RT-PCR?
We have learned above that RT-PCR Full Form stands for Reverse Transcription-Polymerase Chain Reaction. It is a nuclear-derived technique used to identify a specific genetic component in any infection or virus.
Originally, markers of radioactive isotopes were used to locate the desired genetic components, which were targeted. But nowadays, isotopic labelling has been replaced with special markers using fluorescence dyes in refining. This technology helps scientists by showing quick outcomes in the middle of a process or even when the process is not yet complete. However, if we employ the traditional RT-PCR technique. The outcome is then only visible during the last procedure.
This real-time RT-PCR technique is useful in a variety of situations. The current reason for finding the COVID-19 virus in the human body can be considered. These techniques are employed in many countries to find other diseases. The COVID-19 virus, Ebola virus and others serve as examples. Additionally, it contributes to increasing testing across the country.
What is a Virus?
A virus is a micro, contagious organism that can only grow inside of living cells. Virus infections are possible in all living things, including plants, animals, and microorganisms like bacteria and archaea. The virus is a kind of biological entity. In a cell, intracellular parasites bind by a Virus. They need to infect living cells in order to reproduce because they are unable to do so on their own. By comparison with the word “bacteria,” which refers to “a rod-like bacterium,” the term “virus” was created.
What is DNA?
The full form of DNA is Deoxyribonucleic Acid. DNA is the genetic material in a cell that contains the instructions for creating and maintaining life. DNA is found in the nucleus of all cells and mitochondria. Mitochondria generate energy in the cell. The DNA sequence of the genome is used to identify an individual.
This information is useful for proving someone’s relationship with anyone such as a daughter, son, mother, father and relative in a family etc. Law enforcement agencies use this information to identify criminals and this allows them to identify criminals or victims of crime or disaster by forensic scientists.
What is RNA?
The full form of RNA is Ribonucleic Acid. All living cells contain nucleic acids and RNA contains nucleic acids. RNA plays an important role in the representation of genetic information from DNA to proteins. RNA is a prototype in a cell that produces proteins in a cell. RNA is made up of DNA and has a complementary base sequence to DNA. RNA is an important key in gene exchange, such as mRNA (messenger RNA) production from deoxyribonucleic acid (DNA) and miRNA (microRNA) production from ribosomes in the cell. Reverse transcription of RNA results in cDNA, which is amplified by PCR and detected in RT-PCR.
What is the Real-Time use of Reverse Transcription-Polymerase Chain Reaction?
RT-PCR (reverse transcription-polymerase chain reaction) is a technique used to make multiple copies of DNA or RNA. This technology process produces a single copy of the actual sequence and involves enzymes and nucleotides that carry out this process. And this process itself is called reverse transcription. It begins as RNA and is then converted by a reverse transcriptase enzyme into DNA. Then, using a primer and the proper enzyme, a polymerase chain reaction is used to amplify this DNA strand.
How to know about Covid by RT-PCR technique?
A method called RT-PCR is used to determine whether RNA and DNA are present in a sample. It is used to find out whether the sample contains bacteria, viruses, or other infectious agents. An RNA virus that can infect people such as the COVID-19 virus. This virus can be detected by RT-PCR technique. To perform this reverse transcriptase, an enzyme is used to convert viral RNA into DNA. A polymerase chain reaction or PCR can be used to find this DNA.
What is the process for RT-PCR?
We describe the RT-PCR technique in a process below:
- In this process, the first step is to warm the sample to a temperature of 203 Fahrenheit (95 degrees Celsius)
- This will activate the needed enzymes for RT-PCR
- When the enzymes are activated, we need to extract the RNA from the sample
- This is accomplished by combining MgCl and EDTA with a solution, and then centrifuging the mixture
- And then the supernatant needs to be removed
- Finally, the RNA then precipitates in a bath of liquid nitrogen
- You will receive a number of products from this procedure, including the desired PCR product, total RNA, and cDNA
What are the Advantages of RT-PCR?
- Diagnostic: Both viral and genetic diseases can be diagnosed using RT-PCR as a diagnostic method.
- Efficiency: RT-PCR is relatively a fast and efficient method of detecting Viruses and diseases with nucleic acid.
- Simplicity: RT-PCR is a simple technique or reaction for getting multiple copies of DNA.
- Specificity: Additionally, it provides specificity by amplifying a particular DNA sequence under regulated temperature settings.
- Sensitivity: This method is sensitive since it can provide the necessary DNA sample sequences.
What are the Disadvantages of RT-PCR?
RT-PCR has some disadvantages despite its significant benefits-
- Due to the difficulties in preserving linearity, the exponential expansion of the reverse transcribed complementary DNA (cDNA) during the numerous cycles of PCR results in erroneous end point quantification. The fact that prior knowledge is required to deal with impending consequences is one of the main shortcomings of this RT-PCR. RT-PCR is prone to errors, just as other error-prone systems.
- The other drawback is that results can be skewed even when only a tiny amount of DNA in RNA is present during testing.
- Therefore, researchers opt to prepare separate reagent rooms for RNA analysis to avoid such false results. Humic acids from environmental samples can sometimes give false conclusions. It depends on how clean the lab is overall.
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