Mix primers, designed in the Samplix design tool to amplify a small, ~150 bp, easy-to-amplify region within or adjacent to your 15-100 kb target region, with template DNA, and droplet PCR mix containing enzyme, buffer, and nucleotides
DNA is partitioned into millions of highly stable double emulsion droplets. Primers anneal and amplify small PCR-product for detection of positive droplets containing target region
After amplification, only droplets containing region of interest are fluorescent
Sorting and collection of positive fluorescent droplets, containing target region
DNA from positive droplets containing target region is released. The enriched target DNA is mixed with reagents for amplification step
The DNA target molecules are partitioned in thousands of independent single emulsion droplets
Target DNA is amplified by droplet-based multiple displacement amplification (dMDA). Traditional MDA-bias is eliminated by the partitioning of DNA
Amplified target DNA, is released from droplets and DNA is ready for downstream analysis by long read sequencing, NGS, PCR, microarray etc.
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