RainDrop Digital PCR
The RainDrop Digital PCR shifts the PCR paradigm from a single color per marker approach to a more scalable and precise multi-color and intensity-per-marker method. This novel approach increases sensitivity by generating between 1 million and 10 million pico-liter sized droplets per lane, which is a 500 – 10,000 times improvement over existing PCR methods. Since each droplet encapsulates a single molecule, researchers can quickly determine the absolute number of droplets containing specific target DNA and compare that to the amount of droplets with normal, background wild-type DNA.
Applications:
- Rare event detection, fluid (plasma, cerebral spinal fluid & urine) biopsy, needle in a haystack BCR-ABL, T790M, KRAS, EGFR, BRAF, etc.
- Copy Number Variation
- One step RT-pdPCR (pico-droplet PCR)
- High precision Chromosomal Copy Number, Trisomy, Non-Invasive Prenatal Testing
- miRNA Methylated DNA Bio-markers
- Pre-test and Validation tool for NGS
Sensitivity: To assess the lower limit of detection (LLOD) of an EGFR mutation, varying concentrations of mutant were diluted into genomic wild-type DNA. The assay delivered a linear response (R2 = 0.998) down to 1 mutant amongst 250,000 wild-type molecules, with a LLOD of 1 in more than 1,000,000 as defined by the average of the wild-type only controls plus 3x the standard deviation.
Multiplexing: By adjusting concentrations of the probes for each individual assay, 4 mutations plus wild-type were measured simultaneously with just VIC and FAM fluorophores. Multiplexing can be expanded to higher plex levels, and it enables detection, identification, and measurement of multiple possible mutations from a single DNA sample