Microfabricated droplet generator for single molecule/cell genetic analysis
Uniform and controlled picoliter or nanoliter-volume droplets (e.g., droplet size varying no more than a few percent)Controlled generation frequencyEffective target incorporation
Genetic and gene expression analyses, combining the advantages of efficient single-molecule amplification in picoliter or nanoliter-volume engineered emulsions and effective product transfer using primer functionalized bead encapsulation.
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Patent Number: US8454906B2
Application Number: US2010670377A
Inventor: Mathies, Richard A. | Kumaresan, Palani | Yang, Chaoyang | Blazej, Robert G.
Priority Date: 24 Jul 2007
Priority Number: US8454906B2
Application Date: 11 May 2010
Publication Date: 4 Jun 2013
IPC Current: B01L000300 | B01L000302 | G01N000110
US Class: 422505 | 422417 | 422501 | 422502 | 422503 | 422504 | 422515 | 422521 | 436180
Assignee Applicant: The Regents of the University of California
Title: Microfabricated droplet generator for single molecule/cell genetic analysis in engineered monodispersed emulsions
Usefulness: Microfabricated droplet generator for single molecule/cell genetic analysis in engineered monodispersed emulsions
Summary: For single molecule/cell genetic analysis in engineered, monodispersed emulsion.
Novelty: Microdroplet generator useful for single cell genetic analysis comprises reagent inlet to reagent channel; oil inlet to oil channel; pneumatic channel; first fluidic layer; elastomeric layer; three-valve pump; and droplet outlet
Chemical/Material
Chemical/Material Application
8454906
Brief Description Improvements in labeling and separation methods coupled with many improvements in the other aspects of Sanger sequencing, facilitated the sequencing of the human genome. However, this cost and production process is not sufficiently cheap or efficient to enable the routine sequencing or re-sequencing of a mammalian genome. Conventional methods of PCR amplification do not allow single cell amplification to produce a uniform amount of template. Current "shake and bake" methods of generating emulsion droplets containing reactants (e.g., via agitation) produce polydisperse droplets, having a wide range of sizes and containing widely varying amounts of target and reagent. UC Scientists have described microfluidic designs and methods for rapid generation of engineered, monodisperse (i.e., uniform-size) picoliter to nanoliter volume droplets of reagent/target (molecule or cell) mix in emulsion oil. The designs and methods enable high-throughput encapsulation of a single target (e.g., DNA/RNA molecules or cells) in controlled size droplets of reagent mix. The pulsatile flow profile of the microfabricated pump provides active control over droplet generation, thereby enabling droplet formation with oils that are compatible with biological reactions but are otherwise difficult to form emulsions with. Publications High-Throughput Single Copy DNA Amplification and Cell Analysis in Engineered Nanoliter Droplets Tech ID/UC Case 17844/2007-124-0 Related Cases 2007-124-0
USA
