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ENIGMA

Ecosystems and Networks Integrated with Genes and Molecular Assemblies

Digital Droplet Multiple Displacement Amplification


 

DigitalDropletDisplacementAmplification_Singh
Fig. 1. The working principles of ddMDA. The ddMDA procedure partitions E.coli sample into millions of picoliter droplets(A) and amplification in each droplet occur independently from the other droplets(B). At ddMDA endpoint, each droplet contains a discrete hyper-branched MDA product(C). Scale bar =100µm. Fig. 2. Comparison of whole-genome coverage of assembled contigs mapped to E. coli K-12 genome for ddMDA and tube MDA. (A) From the outermost circle, ddMDA for 100 pg/µL, tube MDA for 100 pg/µL, 10 pg/µL, and 10 pg/µL and (B) ddMDA for 1 pg/µL, tube MDA for 1 pg/µL, 0.1 pg/µL, and 0.1 pg/µL, respectively. GC contents in black and genomic DNA in green as a reference.
  • The ddMDA technique enabled significantly lower bias and non-specific amplification than conventional MDA thus achieving more uniform coverage of amplification over the entire genome
  • This technique can be a powerful tool for genomic studies where DNA samples are limited such as single cells, microaggregates, and uncultured microbes from many different environments

Digital Droplet Multiple Displacement Amplification (ddMDA) for whole-genome sequencing of limited DNA samples (2016) Rhee, M., Light, Y.K., Meagher, R.J., and Singh, A.K. Plos One [doi]10.1371/journal.pone.0153699