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Manoj, P (2016) Droplet digital PCR technology promises new applications and research areas. Mitochondrial DNA. Part A, DNA mapping, sequencing, and analysis, 27 (1). pp. 742-6. ISSN 2470-1408

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Abstract

Digital Polymerase Chain Reaction (dPCR) is used to quantify nucleic acids and its applications are in the detection and precise quantification of low-level pathogens, rare genetic sequences, quantification of copy number variants, rare mutations and in relative gene expressions. Here the PCR is performed in large number of reaction chambers or partitions and the reaction is carried out in each partition individually. This separation allows a more reliable collection and sensitive measurement of nucleic acid. Results are calculated by counting amplified target sequence (positive droplets) and the number of partitions in which there is no amplification (negative droplets). The mean number of target sequences was calculated by Poisson Algorithm. Poisson correction compensates the presence of more than one copy of target gene in any droplets. The method provides information with accuracy and precision which is highly reproducible and less susceptible to inhibitors than qPCR. It has been demonstrated in studying variations in gene sequences, such as copy number variants and point mutations, distinguishing differences between expression of nearly identical alleles, assessment of clinically relevant genetic variations and it is routinely used for clonal amplification of samples for NGS methods. dPCR enables more reliable predictors of tumor status and patient prognosis by absolute quantitation using reference normalizations. Rare mitochondrial DNA deletions associated with a range of diseases and disorders as well as aging can be accurately detected with droplet digital PCR.

Item Type: Article
Uncontrolled Keywords: Gene copy number, qPCR, real-time PCR
Subjects: Human Molecular Genetics
Depositing User: Rgcb Library
Date Deposited: 30 Mar 2017 08:40
Last Modified: 12 Apr 2019 08:06
URI: http://rgcb.sciencecentral.in/id/eprint/334

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