Abstract: Some examples herein provide a sequencing flowcell that includes an imaging sensor, a hydrogel disposed on the imaging sensor and comprising a moiety; and a first complex non-covalently coupled to the moiety, the first complex comprising a first oligonucleotide. A method of using the sequencing flowcell may include decoupling the first complex from the moiety; and coupling a second complex to the moiety, the second complex comprising a second oligonucleotide. Methods of forming the flowcell are also provided.

Abstract: 3?-blocked nucleotides, methods of deblocking the same, and methods of synthesizing polynucleotides using the same are provided herein. In some examples, a nucleotide is disposed within the aperture on the first side of a nanopore. The nucleotide may be coupled to a 3?-blocking group including a trigger. The trigger may be selectively activated using an initiator. The activated trigger may be used to remove the 3?-blocking group from the nucleotide.

Abstract: In one example, a method of preparing a fluidic channel includes covalently coupling a first region of a substrate to a first region of a cover using first moieties covalently coupled to the first region of the substrate and second moieties covalently coupled to the first region of the cover. The covalent coupling between the first region of the substrate and the first region of the cover suspends a second region of the cover over a second region of the substrate to form a fluidic channel.

Abstract: The invention relates to deformable polymers comprising immobilised primers, particularly for use in nucleic acid sequencing, such as concurrent sequencing.

Abstract: Some examples herein provide a sequencing flowcell that includes an imaging sensor; a hydrogel disposed on the imaging sensor and comprising a moiety; and a first complex non-covalently coupled to the moiety, the first complex comprising a first oligonucleotide. A method of using the sequencing flowcell may include decoupling the first complex from the moiety; and coupling a second complex to the moiety, the second complex comprising a second oligonucleotide. Methods of forming the flowcell are also provided.

Abstract: In some examples, a method of capturing a polynucleotide on a particle that includes a capture primer includes transporting the particle to a first aperture between a first fluidic compartment and a second fluidic compartment. The particle may be located in the first fluidic compartment, and the polynucleotide may be at least partially located in the second fluidic compartment. The method may include transporting the polynucleotide from the second fluidic compartment to the first fluidic compartment through the first aperture. The method may include hybridizing the polynucleotide to the capture primer.

Abstract: Some examples herein provide a method that includes reacting an unsaturated cyclic dione with an indole or indazole to form a first adduct. One of a substrate and a functional group is coupled to the unsaturated cyclic dione, and the other of the substrate and the functional group is coupled to the indole or indazole. Forming the first adduct couples the functional group to the substrate. Some examples herein provide a composition that includes a substate, a functional group, and an adduct coupling the functional group to the substrate. The adduct may be a product of a reaction between an unsaturated cyclic dione and an indole or indazole.

Abstract: In some examples, a structure is contacted with polynucleotides having a variety of lengths and each including first and second adapters. The structure includes a substrate including first and second regions spaced apart from one another by a gap of at least 100 nm, a first set of capture primers coupled to the first region of the substrate, and a second set of capture primers coupled to the second region of the substrate. The first adapter of the polynucleotide is hybridized to a capture primer of the first set of capture primers. Based on that polynucleotide being sufficiently long to bridge the gap, it is amplified using the first and second sets of capture primers. Based upon that polynucleotide being insufficiently long to bridge the gap, it is not amplified. Optionally, a wall may be disposed in the gap.

Abstract: In some examples, a method includes disposing a first hydrogel within a first recess of a substrate and over a pillar. The substrate may include a second recess in which the pillar is disposed, and a wall separating the first recess from the second recess. While the first hydrogel is disposed within the first recess, the pillar may be removed. After removing the pillar, a second hydrogel may be disposed within the second recess. Also provided herein are nonlimiting examples of manners in which recesses, walls, and patterned hydrogels may be formed.

Abstract: Automated methods conducted in a sequencing flowcell, and kits for reusing a flowcell, are provided herein. In some examples, an automated method conducted in a sequencing flowcell may include, at a surface of the sequencing flowcell coupled to a first moiety, using a reagent to decouple a first complex from the first moiety. In some examples, the first complex may include a second moiety which couples to the first moiety and a polynucleotide coupled to the second moiety. In some examples, the method may further include using a nuclease to polynucleotides in the sequencing flowcell. The method may include, after using the reagent and after using the nuclease, coupling a second complex to the first moiety. The second complex may include a third moiety which couples with the first moiety and an oligonucleotide coupled to the third moiety.

Abstract: In one example, an unsaturated cyclic dione is coupled to the substrate, and is reacted with an indole or indazole including a first functional group to form a first adduct coupling the first functional group to the substrate. In another example, an unsaturated cyclic dione is coupled to a substrate and reacted with a diene including a functional group to form an adduct coupling the functional group to the substrate. In another example, an indole or indazole is coupled to a substrate, and is reacted with an unsaturated cyclic dione including an oligonucleotide to form an adduct coupling the oligonucleotide to the substrate. In another example, a diene is coupled to a substrate, and is reacted with an unsaturated cyclic dione including an oligonucleotide to form an adduct coupling the oligonucleotide to the substrate.

Abstract: In some examples, novel photochemically-reversible hydrogels and nanogel particles are described comprising copolymer chains including at least one reactive alkene or reactive 1,4-diene end group capable of [2+2] or [2+2+2+2] photodimerization, respectively, at wavelengths >270 nm. In various examples, the photochemically-reversible hydrogels comprise copolymer chains including at least one —N3, —C?CH or —CO2H end group for dual functionality and/or pH responsiveness. For nucleic acid sequencing, amplification primers are grafted to photochemically-reversible hydrogels or nanogel particles reversibly bound to surfaces within a flow cell. After sequencing is complete, the photochemically-reversible hydrogel or nanogel particles is/are removable from the flow cell surfaces by irradiation, enabling the flow cell to be reusable.

Abstract: Reusable flow cells for sequencing which exhibit signal intensity retention over numerous use cycles, the active surface of which contains poly-azide functional moieties, methods of treating flow cells surfaces with reagents to provide such poly-azide functional moieties, and reagents therefor.

Abstract: In some examples, novel nanogel particles are described having dual functionality, temperature responsiveness and pH responsiveness. For nucleic acid sequencing, amplification primers are grafted to nanogel particles to form primer-grafted nanogel particles, and the primer-grafted nanogel particles are captured onto surfaces within a flow cell. Within flow cells such as used in SBS nucleic acid sequencing, each primer-grafted nanogel particle functions as a nano-well in the flow cell, thus eliminating the need for nano-wells in some examples.

Abstract: In one example, a method of preparing a fluidic channel includes covalently coupling a first region of a substrate to a first region of a cover using first moieties covalently coupled to the first region of the substrate and second moieties covalently coupled to the first region of the cover. The covalent coupling between the first region of the substrate and the first region of the cover suspends a second region of the cover over a second region of the substrate to form a fluidic channel.

Abstract: In one example, an unsaturated cyclic dione is coupled to the substrate, and is reacted with an indole or indazole including a first functional group to form a first adduct coupling the first functional group to the substrate. In another example, an unsaturated cyclic dione is coupled to a substrate and reacted with a diene including a functional group to form an adduct coupling the functional group to the substrate. In another example, an indole or indazole is coupled to a substrate, and is reacted with an unsaturated cyclic dione including an oligonucleotide to form an adduct coupling the oligonucleotide to the substrate. In another example, a diene is coupled to a substrate, and is reacted with an unsaturated cyclic dione including an oligonucleotide to form an adduct coupling the oligonucleotide to the substrate.