As far as the polymerase chain reaction (PCR) is concerned, it is one of the robust technologies employed in molecular biology. The process helps in duplicating or increasing particular DNA sequence or any complementary template using oligonucleotides, thermal cycling, and heat-stable DNA polymerase. With every cycle, the amount of target molecules is doubled in a theoretical manner with the DNA exponentially improved. According to an article published on https://www.ncbi.nlm.nih.gov, several molecular techniques such as hybridization assay, biosensor detection, sandwich hybridization assay, and real-time PCR assay have been built. These methods are used for precise and particular recognition and assessment of marine toxic microalgae species. To learn more about real-time PCR, keep reading this article.
Benefits of real-time PCR
It is true that standard and real-time PCR have a similar process, there are many benefits of the latter. Here are the major ones:
Identify enlarged pieces: The first and foremost benefit is that real-time PCR helps in the identification of the augmented fragments during the process. The method gauges the product amount during the exponential stage. The standard PCR method, on the contrary, determines the amount of the product during the plateau stage. It is more beneficial to gauge during the exponential stage. That is because the measurements noted during the plateau stage fail to show the amount of the starting material.
Standard PCR is time-consuming: When it comes to standard PCR, it calls for post-PCR assessment and recognizes product through sequence or size. Though using gel electrophoresis is not costly, the process takes time compared to realtime PCRvs PCR. Moreover, the standard PCR is not automated. Besides, the process scores low concerning specificity because the molecules of similar weights are hard to identify. As far as gel electrophoresis is concerned, it is not asuitable option for endpoint evaluation or assessment for most of the lab processes or techniques.
The employment of probe hybridization is frequently employed for product characterization by sequence. The process is informative and dependable, but also time-consuming and costly. When it comes to ELISA detections, they too take much time. As far as real-time PCR is concerned, these problems are eliminated. When it comes to Amplicon identification, it is attained by keeping an eye on the accretion of particular products during every cycle.
The process performed in the same tube: The complete process right from amplification to assessment is done in the same tube. It differs from the standard process where the PCR item is shifted and influenced into different formats. Consequently, there is less possibility of polluting the product with real-time PCR techniques.
The applications include microRNA and non-coding RNA assessment, gene expression or mRNA evaluation, protein analysis, genetic variation analysis, and mutation detection assays.
Now that you know about real-time PCR, it is time that you study more about it in detail. The process helps to monitor individual responses as they occur in real time. It helps in measuring the number of Amplicon in every cycle. The process of detection is dynamic. If you have any more questions about real-time PCR, feel free to ask or comment.