Digital PCR is a PCR method that involves a sensitive or reproducible way of measuring the amount of DNA and RNA present in a sample. Similar to qPCR, the reaction to the components and amplification process is the same, but the difference lies in the way the target of the sample is measured. It is a simple method that does not rely on a calibration curve, and also it does not need any reference standards or other kinds of endogenous controls.
During the initial stage of digital PCR, the sample mix is divided into s large number of individual wells that result in either 1 or 0 targets. However, following the PCR amplification, the number of positive and negative reactions is determined, and the quantification of the target is calculated by using Poisson statistics. This process is also a powerful method that has a number of advantages like it has improved rare allele and CNV detection increased sensitivity for detecting the clinical samples, etc.
Also, the system provides a precise measure of DNA molecules in each drop. Because the precision is given within single wells and can be multiplied by using multiple wells, this process can be used incredibly for cancer research. There are many sites that offer more knowledge about this system so you can research more on this website and find out more about Digital PCR.
Things you need to know about digital PCR
Here are four things you need to know about Digital PCR: –
- There are several advantages of digital PCR, and this is the reason it is used instead of real-time quantitative PCR in clinical microbiology. The dPCR method is a lot dependent on the use of a calibration curve and is less affected by suboptimal amplification. Also, there are some studies that have shown improved accuracy and precision based on target polymorphism s or differences in assay design. Also, viral quantitation is an attractive application as the viral load assays have become an important part of diagnosing the diseases and prediction of the clinical outcome. Also, it helps to monitor the therapeutic response and the initiation of preemptive treatment. The accurate quantitation and viral reservoirs, and because of this, it is also used in the treatment of HIV. Apart from this, Digital PCR can also help to reduce the variability in the laboratories and use it as a reference against which other control materials can be measured.
- Digital PCR is also considered as a novel method that is used for precise quantification of the nucleic acids. In this method, similar assay reagents are used that are also used in standard analog measurements. However, the counts of the total number of individual target molecules are presented in a digital format that also enables many functions that require high sensitivity. Also, it is used in a clinical setting because absolute concentrations are easily determined. And the data can be easily interpreted and compared across different samples.
- It is also used for other purposes like mutation detection and copying a number of analyses on cell-free DNA and the DNA extraction from FFPE tissue in case of a cancer diagnosis. Also, it is utilized for the quantification of the pathogenic organisms with splice variant quantification. However, the quantification of reference materials is used in GMO analysis. Additionally, it is also used in gene copy number quantification in order to stabilize transgenic cells or organisms and analysis of the biodistribution of CAR T-cells in the preclinical models. It is also used for various other customized dPCR based applications. They are also used for other purposes, and their uses are cited in various websites, and if interested, you can access the information on this website.
- It is also used for higher accuracy, sensitivity, and absolute quantification as it is a new approach to conventional quantitative PCR for rare allele detection. In this process, a single molecule can be amplified to a million fold or more, and during the amplification process, dye-labeled probes are used in order to detect sequence-specific targets. However, when there is no sequence target, there is no accumulation of signals. The PCR analysis also involves negative reactions, and it is used to generate an absolute count of a number of target molecules without the need for endogenous controls.
It can be seen that PCR and its variant has been revolutionized the practice of clinical microbiology. And with the continuous advancements in PCR have led to new technology, i.e., Digital PCR, which has a number of advantages. It offers many benefits as compared to the traditional PCR, and because of this reason, many laboratories are considering the implementation of dPCR, which uses a powerful technique for specific applications. There are many websites that offer knowledge about its order to know more about digital PCR; you can access the information on this website.