DROPLET-BASED MICROFLUIDIC PLATFORMS FOR HIGH-THROUGHPUT DNA GENOTYPING AND AMPLIFICATION

microfluidics;genotyping;Biomedical Engineering

Zheng, TonyWang, Jeff T. ;

Johns Hopkins University

关键词: microfluidics;    genotyping;    Biomedical Engineering;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/44647/ZHENG-THESIS-2017.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

Common genetic analysis instruments in genotyping, allele discrimination, and PCR are standardized, indispensable tools in biological, medical, and science research laboratories and R&D industry sectors across the world. Constantly working to improve and create new applications, many fields are looking to construct next-generation technologies that can offer higher throughput, higher efficiency, and lower costs in such genetic analysis techniques. In particular, multinational agricultural companies, in the face of a rapidly growing human population, are working to improve crop production and to accelerate plant breeding and genetic selection techniques in order to maintain global food supplies. These companies, such as DuPont Pioneer we are in collaboration with, are now turning to new microfluidic technologies that present a promising route for miniaturizing and up scaling high-throughput genotyping. Microfluidic technologies reduce sample consumption by thousands, up to millions fold from milliliter volumes to nanoliter droplets, lift processing constraints from discrete 96-well plates to an unlimited, continuous-flow of droplet processing, and downsize of physical footprint from bulky, benchtop instruments to palm-sized, miniaturized devices.Here, we present two novel microfluidic platforms: 1) a microfluidic platform for multiplexed genotyping of single nucleotide polymorphisms (SNPs), screening a series of DNA targets against a library of probes, achieving 100% accuracy in SNP calling of synthetic maize DNA targets and 93% accuracy in SNP calling of genomic maize DNA targets, using an Invader assay, and 2) a microfluidic platform for integrated, continuous-flow, droplet Taqman-based PCR and end-point allele discrimination. The development of both microfluidic devices offers promising platforms for next generation, high throughput genotyping technologies.