Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
droplet Digital PCR
Droplet Digital PCR (ddPCR) is a sensitive and quantitative method for detection of any nucleotide-target sequences in biological and environmental samples with multiplexing capability.
The ddPCR method is based on compartmentalizing a PCR reaction into thousands of droplets, by which the target sequences get randomly distributed between these nano reactions. Using target-specific fluorescent probes within the PCR reaction for detection, droplets containing the target-sequence will turn fluorescent, while other droplets stay negative. The distribution of positive (=1) and negative droplets (=0) is subsequently analyzed with a ddPCR analyzer, translating the sample into a digital readout based on ones and zeros.
Since the distribution of target sequences into droplets follows the Poisson law, an absolute quantification of target sequences present in the sample can be performed using Poisson Distribution Analysis. As this eliminates the need for standard curves and avoids reliance on arbitrary expression units, ddPCR has a major advantage as compared to conventional qPCR methods, especially if an absolute quantification is desirable.
Using Biorads QX600 analyzer with 6 fluorescent channels also allows multiplexing, making it a time and cost-efficient method. While 6 target sequences can easily be analyzed at a time, the system can be tweaked further for detection of 12 targets following optimization.
Hence ddPCR is a versatile tool that can be used for a broad range of applications such as:
