Droplet Digital Polymerase Chain Reaction: A Novel Method for Absolute Quantifi cation of the Target DNA
Year: 2013
Type: article
Abstract: The polymerase chain reaction (PCR) is a method for amplifying specifi c segments of the target DNA under controlled conditions.Under the concept of no use of the standard, digital PCR (dPCR) was developed for absolute quantifi cation of the target DNA by using endpoint measurement.On the basis of the Poisson distribution statistic of dPCR, the DNA target was randomly distributed into an individual partition, it then employs the signal detection of the positive or negative partitions to calculate the amount of the target DNA.The droplet digital PCR (ddPCR) is a novel type of dPCR, which bases on generating the DNA into water-in-oil droplets.The ddPCR requires Taqman assay as the detection method similarly to the real time PCR, to provide a fl uorescence signal for determination.The working process of ddPCR runs from generating the emulsion droplets, thermal cycling droplets in a normal thermal cycler till the endpoint, and analyzing the data by using a droplet reader assembled with fl uorescent detection machine.The applications of ddPCR nowadays run into 3 areas including analysis of copy number of variations (CNVs), absolute quantifi cation of biomarker DNA in a cell-free plasma sample, and detection of rare allele with small fold of target differences such as mutation gene in a wild type DNA.
Authors: Nattharat Jearapong, Kanokwan Jarukamjorn
Language: Thai
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W1488200546