As demands to maintain the global food production continue to mount,multinational seed companies are turning to new DNA marker technologies to accelerate the rate of plant breeding and crop improvement.The key to wide...As demands to maintain the global food production continue to mount,multinational seed companies are turning to new DNA marker technologies to accelerate the rate of plant breeding and crop improvement.The key to widespread adoption of molecular breeding is the availability of flexible and cost-effective tools that can perform combinatorial and high-throughput genotyping of single-nucleotide polymorphisms(SNPs)to guide the crop development process.Toward this end,we have developed a programmable,droplet-based microfluidic device for genotyping maize genomic DNA.A unique feature of the microfluidic platform is the nano sample processors(NSPs),which allow the device to sequentially load an unrestricted number of unique DNA samples using only two inlets,overcoming the current limitation to the number of sample inputs due to small device footprint.Direct and programmable droplet generation within the device allows each sample to be genotyped against a panel of markers on demand.Moreover,we have successfully implemented the Invader assay for SNP genotyping in flowing,50-nL droplets,thus achieving significant reduction in consumption of reagents per reaction as compared with conventional genotyping platforms.As a demonstration,we performed 240 Invader reactions(testing 8 DNA samples against 10 SNP markers)and achieved greater than 93% accuracy in SNP calling of plant DNA samples in a single droplet-based experiment.展开更多
基金We thank the funding support from Dupont Pioneer and NSF(Grant No.1538813).
文摘As demands to maintain the global food production continue to mount,multinational seed companies are turning to new DNA marker technologies to accelerate the rate of plant breeding and crop improvement.The key to widespread adoption of molecular breeding is the availability of flexible and cost-effective tools that can perform combinatorial and high-throughput genotyping of single-nucleotide polymorphisms(SNPs)to guide the crop development process.Toward this end,we have developed a programmable,droplet-based microfluidic device for genotyping maize genomic DNA.A unique feature of the microfluidic platform is the nano sample processors(NSPs),which allow the device to sequentially load an unrestricted number of unique DNA samples using only two inlets,overcoming the current limitation to the number of sample inputs due to small device footprint.Direct and programmable droplet generation within the device allows each sample to be genotyped against a panel of markers on demand.Moreover,we have successfully implemented the Invader assay for SNP genotyping in flowing,50-nL droplets,thus achieving significant reduction in consumption of reagents per reaction as compared with conventional genotyping platforms.As a demonstration,we performed 240 Invader reactions(testing 8 DNA samples against 10 SNP markers)and achieved greater than 93% accuracy in SNP calling of plant DNA samples in a single droplet-based experiment.
