Digital PCR: The Emergence of the Digital Age
The Future for PCR Diagnostics
PCR is polymerase chain reaction is in vitro amplification of any given DNA or RNA sample, while digital PCR is the quantitative PCR technique that provide reproducible way of measuring amount of DNA or RNA present in particular sample. The advantage of digital PCR has high tolerance to inhibitors and it distinguish expression of alleles, and measure the cancer genes. PCR has various application such as paternity testing, detection of hereditary disease, forensic science, and DNA cloning.
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The key difference between dPCR and traditional PCR lies in the method of measuring nucleic acids amounts, with the former being a more precise method than PCR, though also more prone to error in the hands of inexperienced users.[1] A "digital" measurement quantitatively and discretely measures a certain variable, whereas an “analog” measurement extrapolates certain measurements based on measured patterns.
PCR carries out one reaction per single sample. dPCR also carries out a single reaction within a sample, however the sample is separated into a large number of partitions and the reaction is carried out in each partition individually. This separation allows a more reliable collection and sensitive measurement of nucleic acid amounts. The method has been demonstrated as useful for studying variations in gene sequences — such as copy number variants and point mutations — and it is routinely used for clonal amplification of samples for next-generation sequencing.
The polymerase chain reaction method is used to quantify nucleic acids by amplifying a nucleic acid molecule with the enzyme DNA polymerase. Conventional PCR is based on the theory that amplification is exponential. Therefore, nucleic acids may be quantified by comparing the number of amplification cycles and amount of PCR end-product to those of a reference sample.
Digital PCR has many applications in basic research, clinical diagnostics and environmental testing. Its uses include pathogen detection and digestive health analysis;liquid biopsy for cancer monitoring, organ transplant rejection monitoring and non-invasive prenatal testing for serious genetic abnormalities;copy number variation analysis, single gene expression analysis, rare sequence detection, gene expression profiling and single-cell analysis; the detection of DNA contaminants in bioprocessing, the validation of gene edits and detection of specific methylation changes in DNA as biomarkers of cancer.
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