Digital PCR (dPCR) uses the procedure of end-point PCR but splits the PCR reaction into many single partitions, in which the template is randomly distributed across all available partitions. After PCR, the amplification target is detected by measuring the fluorescence – of either sequence-specific DNA probes or intercalating dyes – in all positive partitions. As the template is distributed randomly, Poisson statistics can be used to calculate the average amount of target DNA per valid, analyzable partition. The total amount of target DNA in all partitions of a well is calculated by multiplying the amount of average target DNA per partition with the number of valid partitions. Calculation of target concentration is determined by referring to the volume in all analyzable partitions, that is, partitions which were filled with reaction mix. The total number of filled partitions is identified by a fluorescent dye present in the reaction mix itself. Absolute quantification by dPCR eliminates the need for standard curves to determine amounts of target DNA in a given sample.

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Digital Droplet PCR

Rajarata University of Sri Lanka


Product Category/ Test Name (Matrix):

Digital Droplet PCR

Sub Category:

QIAcuity® Digital PCR System

Keywords:

PCR, DNA amplification, microdroplet technology, droplet intelligence, Fully automatic



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Under Development Stage

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