Provides a method to obtain enrichment of large DNA molecules for long-read sequencing using FACS or microfluidic partitioning. Furthermore, the present invention relates to a kit comprising a plurality of 5 microfluidic devices and a plurality of fluids configured for use with the system and the method.
The invention provides a fluorosurfactant composition for use in stabilising emulsions, in particular emulsions comprising single or double-emulsion droplets; as well as improved methods for manufacture of the fluorosurfactant; wherein the surfactant comprises a perfluorocarbon chain amide covalently linked to a polymer of ethylene 5 oxide and propylene oxide. The invention also provides methods for single-cell resolution cell killing- and cell secretion-assays based on droplets stabilised by the fluorosurfactant composition.
The present invention relates to a system for sorting particles by characteristics that can be optically detected. The system comprises a microfluidic device that is capable of sorting the particles, and an instrument driving the fluids through the microfluidic device and invoking the sorting events in response to the optical signals emitted by the particles. The present invention also relates to a method for enrichment or isolation of a nucleotide fragment comprising a known nucleotide sequence element and to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for sorting of emulsion droplets.
Provides a method to obtain enrichment of large DNA molecules for long-read sequencing using FACS or microfluidic partitioning. Furthermore, the present invention relates to a kit comprising a plurality of 5 microfluidic devices and a plurality of fluids configured for use with the system and the method.
The invention provides a fluorosurfactant composition for use in stabilising emulsions, in particular emulsions comprising single or double-emulsion droplets; as well as improved methods for manufacture of the fluorosurfactant; wherein the surfactant comprises a perfluorocarbon chain amide covalently linked to a polymer of ethylene 5 oxide and propylene oxide. The invention also provides methods for single-cell resolution cell killing- and cell secretion-assays based on droplets stabilised by the fluorosurfactant composition.
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
C08G 65/00 - Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
C08G 65/26 - Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring from cyclic ethers and other compounds
C08G 65/332 - Polymers modified by chemical after-treatment with organic compounds containing oxygen containing carboxyl groups, or halides or esters thereof
C08G 65/337 - Polymers modified by chemical after-treatment with organic compounds containing other elements
C08L 71/00 - Compositions of polyethers obtained by reactions forming an ether link in the main chainCompositions of derivatives of such polymers
The present invention relates to a system for amplification of polynucleotides from a predefined number of single cells. The system comprise a device (or part) providing the predefined number of single cells, at a previously defined inlet site (or orifice) of a cartridge (microfluidic device), and the cartridge itself. The invention further relates to a method for amplification of polynucleotides from the one or more single cells using the system to provide an emulsion of aqueous droplets wherein the nucleic acid amplification occurs. Furthermore, the present invention relates to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the system and the method.
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
The present invention relates to a system for sorting particles by characteristics that can be optically detected. The system comprises a microfluidic device that is capable of sorting the particles, and an instrument driving the fluids through the microfluidic device and invoking the sorting events in response to the optical signals emitted by the particles. The present invention also relates to a method for enrichment or isolation of a nucleotide fragment comprising a known nucleotide sequence element and to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for sorting of emulsion droplets.
A microfluidic device, a method for manufacturing a microfluidic device, and a method for provision of double emulsion droplets using a microfluidic device. Furthermore, an assembly configured to supply pressure to the microfluidic device for provision of double emulsion droplets. Furthermore, a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for provision of double emulsion droplets.
A microfluidic device (100) comprises an emulsification section (101) comprising one or more emulsification units (170); and a container section (102) comprising one or more groups of containers comprising one group of containers for each emulsification unit; each emulsification unit (170) comprising a fluid conduit network (135) comprising: a plurality of supply conduits comprising a primary supply conduit (103) and a secondary supply conduit (106); a transfer conduit (112); and a first fluid junction (120) providing fluid communication between the primary supply conduit (103), the secondary supply conduit (106), and the transfer conduit (112); each group of containers (103) comprising a plurality of containers comprising an intermediate chamber (174), a collection container (134), and one or more supply containers (131) comprising a secondary supply container, the secondary supply container (131) defining a secondary supply cavity, the secondary supply container (131) comprising a secondary orifice (177) extending from the secondary supply cavity and a primary orifice (176) extending from the secondary supply cavity, the collection container (134) being in fluid communication with the transfer conduit (112) of the corresponding emulsification unit (170) via a collection orifice of the collection container (134), the secondary supply container (131) being in fluid communication with the secondary supply conduit (106) of the corresponding emulsification unit (170) via the secondary orifice (177), the secondary supply container (131) being in fluid communication with the intermediate chamber (174) of the same group of containers (103) via the primary orifice (176), the intermediate chamber (174) being in fluid communication with the first fluid junction (120) of the corresponding emulsification unit (170) via the primary supply conduit (103) of the corresponding emulsification unit (170). Furthermore a method of manufacturing said device and a method for providing emulsion droplets using such a microfluidic device.
The present invention relates to a microfluidic device, a method for manufacturing a microfluidic device, and a method for provision of double emulsion droplets using a microfluidic device. Furthermore, the present invention relates to an assembly configured to supply pressure to the microfluidic device for provision of double emulsion droplets. Furthermore, the present invention relates to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for provision of double emulsion droplets. The microfluidic device comprises a transfer conduit comprising a first transfer conduit part having a first affinity for water; and a collection conduit comprising a first collection conduit part having a second affinity for water being different from the first affinity for water. A well section and a microfluidic section of the microfluidic device are fixedly connected to each other.
B01F 33/3011 - Micromixers using specific means for arranging the streams to be mixed, e.g. channel geometries or dispositions using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
B01F 33/301 - Micromixers using specific means for arranging the streams to be mixed, e.g. channel geometries or dispositions
The present invention relates to a system for amplification of polynucleotides from a predefined number of single cells. The system comprise a device (or part) providing the predefined number of single cells, at a previously defined inlet site (or orifice) of a cartridge (microfluidic device), and the cartridge itself. The invention further relates to a method for amplification of polynucleotides from the one or more single cells using the system to provide an emulsion of aqueous droplets wherein the nucleic acid amplification occurs. Furthermore, the present invention relates to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the system and the method.
The present invention pertains to an in vitro method in which a targeted DNA molecule containing a DNA sequence of interest is enriched by a) general amplification of DNA molecules in a multiple of droplets each containing less than 0.5 target DNA molecule on average (404), b) specific detection of the target DNA molecule in each of the droplets (405), and c) physically selecting droplets containing target DNA molecules (406).
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
The present invention relates to a microfluidic device and method for providing emulsion droplets. The device comprising: a microfluidic section comprising one or more microfluidic units; and a well section comprising one or more groups of wells comprising one group of wells for each microfluidic unit; the well section and the microfluidic section forming a fixedly connected unit such that each group of wells forms a fixedly connected unit with a respective corresponding microfluidic unit, each microfluidic unit comprising a fluid conduit network comprising: a plurality of supply conduits comprising a secondary supply conduit and a primary supply conduit comprising a capillary structure having a volume of at least 2 μL; a transfer conduit; and a first fluid junction providing fluid communication between the primary supply conduit, the secondary supply conduit, and the transfer conduit; each group of wells comprising a plurality of wells comprising a collection well and one or more supply wells comprising a primary supply well, the collection well being in fluid communication with the transfer conduit of the corresponding microfluidic unit, the primary supply well being in fluid communication with the primary supply conduit and the secondary supply conduit of the corresponding microfluidic unit.
B01F 33/3011 - Micromixers using specific means for arranging the streams to be mixed, e.g. channel geometries or dispositions using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
14.
Targeted enrichment of long nucleotide sequences using microfluidic partitioning
The present disclosure relates to methods for enrichment or isolation of a long nucleotide fragment comprising a known nucleotide sequence element, i.e. a sequence encoding a conserved active site or domain, the method being applicable i.a. to high throughput screening for DNA fragments containing a known sequence element. The methods herein comprise the steps of forming an emulsion of multiple liquid droplets from a DNA fragment containing liquid sample, specific detection of droplets containing at least one target DNA molecule, physically selecting droplets containing at least one target DNA molecule, and performing a general amplification of the DNA molecules containing at least one target DNA molecules.
C12P 19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
A microfluidic device, a method for manufacturing a microfluidic device, and a method for provision of double emulsion droplets using a microfluidic device. Furthermore, an assembly configured to supply pressure to the microfluidic device for provision of double emulsion droplets. Furthermore, a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for provision of double emulsion droplets.
A microfluidic device (100) comprises an emulsification section (101) comprising one or more emulsification units (170); and a container section (102) comprising one or more groups of containers comprising one group of containers for each emulsification unit; each emulsification unit (170) comprising a fluid conduit network (135) comprising: a plurality of supply conduits comprising a primary supply conduit (103) and a secondary supply conduit (106); a transfer conduit (112); and a first fluid junction (120) providing fluid communication between the primary supply conduit (103), the secondary supply conduit (106), and the transfer conduit (112); each group of containers (103) comprising a plurality of containers comprising an intermediate chamber (174), a collection container (134), and one or more supply containers (131) comprising a secondary supply container, the secondary supply container (131) defining a secondary supply cavity, the secondary supply container (131) comprising a secondary orifice (177) extending from the secondary supply cavity and a primary orifice (176) extending from the secondary supply cavity, the collection container (134) being in fluid communication with the transfer conduit (112) of the corresponding emulsification unit (170) via a collection orifice of the collection container (134), the secondary supply container (131) being in fluid communication with the secondary supply conduit (106) of the corresponding emulsification unit (170) via the secondary orifice (177), the secondary supply container (131) being in fluid communication with the intermediate chamber (174) of the same group of containers (103) via the primary orifice (176), the intermediate chamber (174) being in fluid communication with the first fluid junction (120) of the corresponding emulsification unit (170) via the primary supply conduit (103) of the corresponding emulsification unit (170). Furthermore a method of manufacturing said device and a method for providing emulsion droplets using such a microfluidic device.
A microfluidic device (100) comprises an emulsification section (101) comprising one or more emulsification units (170); and a container section (102) comprising one or more groups of containers comprising one group of containers for each emulsification unit; each emulsification unit (170) comprising a fluid conduit network (135) comprising: a plurality of supply conduits comprising a primary supply conduit (103) and a secondary supply conduit (106); a transfer conduit (112); and a first fluid junction (120) providing fluid communication between the primary supply conduit (103), the secondary supply conduit (106), and the transfer conduit (112); each group of containers (103) comprising a plurality of containers comprising an intermediate chamber (174), a collection container (134), and one or more supply containers (131) comprising a secondary supply container, the secondary supply container (131) defining a secondary supply cavity, the secondary supply container (131) comprising a secondary orifice (177) extending from the secondary supply cavity and a primary orifice (176) extending from the secondary supply cavity, the collection container (134) being in fluid communication with the transfer conduit (112) of the corresponding emulsification unit (170) via a collection orifice of the collection container (134), the secondary supply container (131) being in fluid communication with the secondary supply conduit (106) of the corresponding emulsification unit (170) via the secondary orifice (177), the secondary supply container (131) being in fluid communication with the intermediate chamber (174) of the same group of containers (103) via the primary orifice (176), the intermediate chamber (174) being in fluid communication with the first fluid junction (120) of the corresponding emulsification unit (170) via the primary supply conduit (103) of the corresponding emulsification unit (170). Furthermore a method of manufacturing said device and a method for providing emulsion droplets using such a microfluidic device.
A microfluidic device, a method for manufacturing a microfluidic device, and a method for provision of double emulsion droplets using a microfluidic device. Furthermore, an assembly configured to supply pressure to the microfluidic device for provision of double emulsion droplets. Furthermore, a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for provision of double emulsion droplets.
The present invention pertains to an in vitro method in which the frequency of the targeted nucleotide sequence containing the DNA fragment of interest is increased stepwise, by several rounds of 1) dilution of a sample containing the DNA fragment of interest into several replicates (separation), 2) randomly amplifying DNA in the replicates (concentration), 3) detecting the DNA fragment of interest in at least one of the diluted and amplified replicates (selection) and repeating steps 1) through 3) until the DNA fragment of interest can be sequenced by standard sequencing techniques.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C12P 19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
01 - Chemical and biological materials for industrial, scientific and agricultural use
05 - Pharmaceutical, veterinary and sanitary products
09 - Scientific and electric apparatus and instruments
10 - Medical apparatus and instruments
42 - Scientific, technological and industrial services, research and design
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
(Based on 44(e)) Chemical preparations and reagents for use in droplet-based single molecule handling; reagents in kit form for use in droplet- based single molecule handling composed of emulsion-forming chemicals, buffers, and reagents to amplify nucleic acid for use in the biotechnology field (Based on 44(e)) Droplet-based pharmaceutical and veterinary preparations, droplet-based sanitary preparations, droplet-based medical preparations, and droplet-based reagents for use in single molecule analysis or single molecular screening of DNA or RNA (Based on 44(e)) Instruments and cartridges for microfluidics-based and droplet-based single molecule handling; data processing equipment and computers for single molecule handling, single molecule analysis or single molecule screening of DNA or RNA (Based on Use in Commerce) Apparatus for DNA testing for medical purposes; apparatus for analyzing substances for medical use (Based on 44(e)) Scientific services and research for others in the field of DNA analysis; medical research services in the field of medical laboratory service; research services relating to health and healthcare, namely, biomedical research services; DNA screening for scientific research purposes; advisory, consultancy and information services relating to the aforesaid (Based on 44(e)) Medical services; DNA screening for medical purposes; medical and health services relating to DNA, genetics and genetic testing; advisory, consultancy and information services relating to the aforesaid
01 - Chemical and biological materials for industrial, scientific and agricultural use
05 - Pharmaceutical, veterinary and sanitary products
09 - Scientific and electric apparatus and instruments
10 - Medical apparatus and instruments
Goods & Services
Chemical preparations for scientific use in analyzing nucleic acids; chemical compositions for scientific and research use in analyzing nucleic acids; materials for use in science, namely, compositions for sample preparations; reagents for scientific use in analyzing nucleic acids; reagent kits consisting primarily of emulsion-forming chemicals, buffers, and reagents to amplify nucleic acid for use in biotechnology fields Diagnostic preparations for medical purposes; reagents for medical purposes; chemical preparations for DNA analysis; none of the aforesaid relating to ophthalmic preparations Scientific apparatus and instruments, namely, droplet generators; for sample preparation of nucleic acids; apparatus for DNA testing for scientific purposes; DNA microarrays Apparatus for DNA testing for medical purposes; medical apparatus for analyzing substances, namely, samples of nucleic acids
22.
A MICROFLUIDIC DEVICE AND A METHOD FOR PROVISION OF EMULSION DROPLETS
The present invention relates to a microfluidic device and method for providing emulsion droplets. The device comprising: a microfluidic section comprising one or more microfluidic units; and a well section comprising one or more groups of wells comprising one group of wells for each microfluidic unit; the well section and the microfluidic section forming a fixedly connected unit such that each group of wells forms a fixedly connected unit with a respective corresponding microfluidic unit, each microfluidic unit comprising a fluid conduit network comprising: a plurality of supply conduits comprising a secondary supply conduit and a primary supply conduit comprising a capillary structure having a volume of at least 2 µL; a transfer conduit; and a first fluid junction providing fluid communication between the primary supply conduit, the secondary supply conduit, and the transfer conduit; each group of wells comprising a plurality of wells comprising a collection well and one or more supply wells comprising a primary supply well, the collection well being in fluid communication with the transfer conduit of the corresponding microfluidic unit, the primary supply well being in fluid communication with the primary supply conduit and the secondary supply conduit of the corresponding microfluidic unit.
The present invention relates to a microfluidic device, a method for manufacturing a microfluidic device, and a method for provision of double emulsion droplets using a microfluidic device. Furthermore, the present invention relates to an assembly configured to supply pressure to the microfluidic device for provision of double emulsion droplets. Furthermore, the present invention relates to a kit comprising a plurality of microfluidic devices and a plurality of fluids configured for use with the microfluidic device for provision of double emulsion droplets. The microfluidic device comprises a transfer conduit comprising a first transfer conduit part having a first affinity for water; and a collection conduit comprising a first collection conduit part having a second affinity for water being different from the first affinity for water. A well section and a microfluidic section of the microfluidic device are fixedly connected to each other.
The invention relates to a method for enrichment or isolation of a long nucleotide fragment comprising a known nucleotide sequence element, i.e. a sequence encoding a conserved active site or domain, the method being applicable i.a. to high throughput screening for DNA fragments containing a known sequence element. The methods include, inter alia, steps of providing a liquid sample of mixed DNA molecules comprising one or more specific target DNA molecules, formation of a multiple of liquid droplets, specific detection of droplets containing at least one of the target DNA molecules, physically selecting droplets containing at least one of the target DNA molecules, and general amplification of the DNA molecules in the coalesced selected droplets.
C12P 19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
01 - Chemical and biological materials for industrial, scientific and agricultural use
05 - Pharmaceutical, veterinary and sanitary products
09 - Scientific and electric apparatus and instruments
10 - Medical apparatus and instruments
Goods & Services
Chemical preparations; chemical compositions and materials for use in science; reagents for scientific purposes; reagents in kit form. Medical preparations; diagnostic preparations and materials; reagents for medical purposes; chemical preparations for DNA analysis; none of the aforesaid relating to ophthalmic preparations. Scientific apparatus and instruments; apparatus for DNA testing for scientific purposes; DNA microarrays. Apparatus for DNA testing for medical purposes; apparatus for analysing substances for medical use.
01 - Chemical and biological materials for industrial, scientific and agricultural use
05 - Pharmaceutical, veterinary and sanitary products
09 - Scientific and electric apparatus and instruments
42 - Scientific, technological and industrial services, research and design
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Chemical preparations, reagents and reagents in kit form for use in droplet-based single molecule handling. Droplet-based pharmaceutical and veterinary preparations, droplet-based sanitary preparations, droplet-based medical preparations, and droplet-based reagents for use in single molecule analysis or single molecular screening of DNA or RNA. Instruments and cartridges for microfluidics-based and droplet-based single molecule handling; data processing equipment and computers for single molecule handling, single molecule analysis or single molecule screening of DNA or RNA. Scientific services and research; medical research services; research services relating to health and healthcare; DNA screening for scientific research purposes; advisory, consultancy and information services relating to the aforesaid. Medical services; DNA screening for medical purposes; medical and health services relating to DNA, genetics and genetic testing; advisory, consultancy and information services relating to the aforesaid.
27.
Nucleotide sequence exclusion enrichment by droplet sorting (NEEDLS)
The present invention pertains to an in vitro method in which a targeted DNA molecule containing a DNA sequence of interest is enriched by a) general amplification of DNA molecules in a multiple of droplets each containing less than 0.5 target DNA molecule on average (404), b) specific detection of the target DNA molecule in each of the droplets (405), and c) physically selecting droplets containing target DNA molecules (406).
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
C12Q 1/6806 - Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
The invention relates to a method for enrichment or isolation of a long nucleotide fragment comprising a known nucleotide sequence element, i.e. a sequence encoding a conserved active site or domain, the method being applicable i.a. to high throughput screening for DNA fragments containing a known sequence element.
The present invention pertains to an in vitro method in which the frequency of the targeted nucleotide sequence containing the DNA fragment of interest is increased stepwise, by several rounds of 1) dilution of a sample containing the DNA fragment of interest into several replicates (separation), 2) randomly amplifying DNA in the replicates (concentration), 3) detecting the DNA fragment of interest in at least one of the diluted and amplified replicates (selection) and repeating steps 1) through 3) until the DNA fragment of interest can be sequenced by standard sequencing techniques.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C12P 19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides
C12Q 1/6848 - Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
The present invention pertains to an in vitro method in which a targeted DNA molecule containing a DNA sequence of interest is enriched by a) general amplification of DNA molecules in a multiple of droplets each containing less than 0.5 target DNA molecule on average(404), b) specific detection of the target DNA molecule in each of the droplets(405), and c) physically selecting droplets containing target DNA molecules (406).
The present invention pertains to an in vitro method in which the frequency of the targeted nucleotide sequence containing the DNA fragment of interest is increased stepwise, by several rounds of 1) dilution of a sample containing the DNA fragment of interest into several replicates (separation), 2) randomly amplifying DNA in the replicates (concentration), 3) detecting the DNA fragment of interest in at least one of the diluted and amplified replicates (selection) and repeating steps 1) through 3) until the DNA fragment of interest can be sequenced by standard sequencing techniques.
