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High-throughput droplet digital pcr system for absolute quantitation

2019-08-19 15:34

Nov 15, 2011 Digital PCR enables the absolute quantitation of nucleic acids in a sample. The lack of scalable and practical technologies for digital PCR implementation has hampered the widespread adoption of this inherently powerful technique. Here we describe a highthroughput droplet digital PCR (ddPCR) systemSample partitioning is the key to digital PCR. In the Droplet Digital PCR (ddPCR) System, a single PCR sample is partitioned into 20, 000 droplets. Template molecules are distributed randomly among droplets, such that some droplets have no template molecules and others have one or more. high-throughput droplet digital pcr system for absolute quantitation

May 25, 2017  Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data Skip to

Here we describe a highthroughput droplet digital PCR (ddPCR) system that enables processing of 2 million PCR reactions using conventional TaqMan assays with a 96well plate workow. Three applications demonstrate that the massive partitioning aorded by our ddPCR system provides orders of magni tude more precision and sensitivity than HighThroughput Droplet Digital PCR System for Absolute Quantitation of DNA Copy Number, B. J. Hindson et Al. ; Vogelstein and Kinzler, Digital PCR. OnChip SingleCopy RealTime ReverseTranscription PCR in Isolated Picoliter Droplets, N. R. Beer et Al. high-throughput droplet digital pcr system for absolute quantitation Available instruments vary with regard to the recommended dynamic range. The BioRad QX100 Droplet Digital PCR system was used to accurately detect rare mutant DNA from 100, 000fold WT 16 and the RainDrop instrument was used to detect a mutated copy from 200, 000 WT copies 13. A typical evaluation of primerprobe sets for use in rare event

HighThroughput Droplet Digital PCR System for Absolute Quantitation of DNA Copy Number Article (PDF Available) in Analytical Chemistry 83(22): November 2011 with 1, 227 Reads high-throughput droplet digital pcr system for absolute quantitation Digital polymerase chain reaction (digital PCR, DigitalPCR, dPCR, or dePCR) is a biotechnological refinement of conventional polymerase chain reaction methods that can be used to directly quantify and clonally amplify nucleic acids strands including DNA, cDNA or RNA. The lack of scalable and practical technologies for digital PCR implementation has hampered the widespread adoption of this inherently powerful technique. Here we describe a highthroughput droplet digital PCR (ddPCR) system that enables processing of 2 million PCR reactions using conventional TaqMan assays with a 96well plate workflow. Droplet Digital PCR (ddPCR) is a method for performing digital PCR that is based on wateroil emulsion droplet technology. A sample is fractionated into 20, 000 droplets, and PCR amplification of the template molecules occurs in each individual droplet. ddPCR technology uses reagents and workflows similar to those used for most standard TaqMan probebased assays.

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