Amplification of nucleic acid, as well as several detection techniques are considered to be the most important tools associated with biological research, currently. Scientists belonging to different areas like basic research, medicine, biotechnology, diagnostics, and forensics are making use of these methods for several applications. In some applications, the detection of qualitative nucleic acid is important. Another application that is in demand is real-time PCR. However, it is important to understand that real-time PCR is being used for both quantitative as well as qualitative analysis. According to research, Real-time PCR can help in diagnosing Zika Virus as well.
What is real-time PCR?
In traditional PCR, the amplified product of DNA, which is also known as an amplicon, is detected within an endpoint analysis. In real-time PCR, this amplification product will be measured as and when there is progress within the reaction, in real-time, with the quantification of the products after every cycle.
- Firstly, the reactions of amplification are going to be set up with the PCR reagents and custom and unique primers.
- After this, the reactions are going to be run within PCR instruments.
- Finally, the data is going to be collected and analyzed with the help of proprietary instrument software.
Real-time detection of several PCR products is normally enabled by including fluorescent reporter molecules in every reaction, which is going to yield increased fluorescence with the increase in the amount of the product DNA. The main benefit associated with real-time PCR is that it helps in determining the total number of template DNA copies with high sensitivity and accuracy. The results of real-time PCR can be qualitative or quantitative whereas PCR is semi-quantitative.
Applications of real-time PCR
Real-time PCR assay has become one of the most important tools for sensitive and rapid determination as well as quantitation of nucleic acid in several biological samples. There are diverse applications, which include the analysis of gene expression, detection of modified organisms within the food, and also, cancer phenotyping.
In the research laboratories, they are used for quantitative measurement of the number of gene copies in transformed cell lines. They can also be used for understanding the changes within gene expression. For instance, the decrease or increase in expression in comparison to the environmental conditions for the treatment of drugs can also be detected by measuring the changes in the cellular levels of mRNA. You can consider purchasing MyBioSource PCR kits directly by visiting the website.
Advantages of real-time PCR
The most important advantage of real-time PCR is that it is highly accurate. The data of real-time PCR can easily be evaluated without the help of gel electrophoresis. This is responsible for reduced bench time as well as increased output. The reactions of real-time PCR help in the evaluation of data in a unified manner. It also helps in reducing contamination. Most importantly, in real-time PCR, there will be an elimination of post-amplification manipulation. This is one important reason why real-time PCR is preferred over PCR.
Conclusion
Real-time PCR is known for the several advantages and unique applications that it has. Make sure that you are going through everything that has been mentioned above to understand why real-time PCR is used as opposed to PCR.
Author Bio: Anthony Karen is a health expert who has been running many health seminars and public discussions. She also manages her blog and reviews the health-related details provided by authentic sources.