The present invention provides methods, compositions, and systems for distributing single polymerase molecules into array regions. In particular, the methods, compositions, and systems of the present invention result in a distribution of single polymerase molecules into array regions at a percentage that is larger than the percentage expected to be occupied under a Poisson distribution.

A method of modifying a microfabricated chip made of a hydrophobic material is provided. The microfabricated chip has a top surface including a plurality of microwells each having a bottom and a side wall, and interstitial space between the microwells. First, the ed chip is surface treated to render the surface of the bottom and side wall of the microwells and the interstitial space hydrophilic. Then, the surface of the interstitial space is treated to render it hydrophobic. A microfabricated chip having hydrophilic microwells and hydrophobic interstitial space is also provided.

The present disclosure is directed to a device (100) comprising a sampling part (110), wherein the sampling part (110) comprises an array of capture zones (112) for sampling liquid volumes between tens of microliters and femtoliters, wherein some or all of the capture zones (112) contain a sponge-like material. Also disclosed are a method for the preparation of such a device and a method for the detection and determination of the presence, concentration and/or properties of an analyte by contacting a liquid sample with such a device.

This invention provides a technique enabling to detect target molecules of low concentration with high sensitivity. This invention includes (i) a step of introducing a hydrophilic solvent (42) containing beads (40),(41) into a space (30) between (a) a lower layer section (10) including a plurality of receptacles (13) each of which is capable of storing only one of the beads (41),(41) and which are separated from each other by a side wall (12) having a hydrophobic upper surface and (b) an upper layer section (20) facing a surface of the lower layer section (10) on which surface the plurality of receptacles (13) are provided; and (ii) a step of introducing a hydrophobic solvent (43) into the space (30), the step (ii) being carried out after the step (i).

An apparatus for biological reactions is provided. The apparatus includes a substrate and a plurality of reaction sites within the substrate. A surface of the substrate is configured to have a first hydrophilicity and each surface of the plurality of reaction sites is configured to have a second hydrophilicity to load a substantial number of reaction sites with a sample volume. The sample volume of each loaded reaction site is substantially confined to its respective reaction site. The sample volume is configured to undergo a biological reaction within the reaction site.

Methods and devices are provided herein for surfaces for de novo nucleic acid synthesis which provide for low error rates. In addition, methods and devices are provided herein for increased nucleic acid mass yield resulting from de novo nucleic acid synthesis.

A miniaturized, automated method for controlled printing of large arrays of nano- to femtoliter droplets by actively transporting mother droplets over hydrophilic-in-hydrophobic (HIH) micropatches. The technology uses single or double-plate devices where mother droplets can be actuated and HIH micropatches on one or both plates of the device where the droplets are printed. Due to the selective wettability of the hydrophilic micropatches in a hydrophobic matrix, large nano- to femtoliter droplet arrays are created when mother droplets are transported over the arrays. The parent droplets are moved by various droplet actuation principles. Also, a method using two plates placed one top another while being separated by a spacer. One plate is dedicated to confirming and guiding parent droplets by using hydrophilic patches in a hydrophobic matrix, while the other plate contains HIH arrays for printing of the droplets. When the parent droplet guidance plate is rotated over the plate dedicated to printing of nano- to femtoliter droplets, the droplets are dispensed inside the HIH array utilizing their selective wettability. The methods allow the parent droplets to move over the HIH arrays many times, providing advantages for performing bio-assays or miniaturized materials synthesis in nano- to femtoliter sized droplets. With controlled evaporation of the dispensed droplets of solution, large arrays of printed material can be generated in seconds. The methods provide a nano- to femtoliter droplet printing technique for a wide variety of applications, e.g., protein- or cell-based bio-assays or printing of crystalline structures, suspensions of nanoparticles or microelectronic components.

De novo synthesized large libraries of nucleic acids are provided herein with low error rates. Further, devices for the manufacturing of high-quality building blocks, such as oligonucleotides, are described herein. Longer nucleic acids can be synthesized in parallel using microfluidic assemblies. Further, methods herein allow for the fast construction of large libraries of long, high-quality genes. Devices for the manufacturing of large libraries of long and high-quality nucleic acids are further described herein.

A method for forming a lipid membrane vesicle includes: filling a chamber with a first aqueous solution by introducing it to a liquid flow path facing a microreactor chip hydrophobic layer main surface; forming a first lipid monolayer membrane in an opening part of the chamber filled with the solution; forming a second lipid monolayer membrane on a layer interface of the organic solvent formed on the main surface of the hydrophobic layer with a second aqueous solution by introducing the solution to the liquid flow path; allowing a first aqueous solution form in the chamber to alter to a spherical droplet covered with the first lipid monolayer membrane; and forming a lipid membrane vesicle by moving the droplet to a position of the second lipid monolayer membrane by applying a physical action, and by zipping the first lipid monolayer membrane covering the droplet and the second lipid monolayer membrane.

A novel miniaturized and highly automated method for the controlled printing of large arrays of nano- to femtoliter droplets is presented by actively transporting mother droplets over hydrophilic-in-hydrophobic micropatches. The proposed technology consists of single plate or double-plate devices where mother droplets can be actuated and hydrophilic-in-hydrophobic micropatches on one or both plates of the device where nano- to femtoliter droplets are printed. Due to the selective wettability of the more wettable hydrophilic micropatches in a hydrophobic matrix, large nano- to femtoliter droplet arrays are created when mother droplets are transported over these arrays. The parent droplets can be moved by different droplet actuation principles, for example, by using the principle of electrowetting-on-dielectric droplet actuation. We propose another method that uses two plates that are placed on top of each other while being separated by a spacer. One plate is dedicated to confirming and guiding of parent droplets by using hydrophilic patches in a hydrophobic matrix, while the other plate contains hydrophilic-in-hydrophobic arrays dedicated to the printing of nano- to femtoliter droplets. When the plate dedicated to parent droplet guiding is rotated over the plate dedicated to printing of nano- to femtoliter droplets, nano- to femtoliter droplets are dispensed inside the hydrophilic-in-hydrophobic array due to their selective wettability. All these proposed methods allow the parent droplets to be moved over the hydrophilic-in-hydrophobic arrays many times, providing unique advantages for performing bio-assays or miniaturized materials synthesis in nano- to femtoliter sized droplets. Upon the controlled evaporation of the dispensed droplets of solution, large arrays of the printed material can be generated on an automated way in seconds of time on a very flexible way. The method disclosed herein provides a distinct nano- to femtoliter droplet printing technique for a wide variety of applications such as protein- or cell-based bio-assays or printing of crystalline structures, suspensions of nanoparticles or components for microelectronics.