A method, in particular an in vitro method, for the circularization of a double stranded DNA nucleic acid. Also, a circularized double stranded DNA nucleic acid obtainable by the method. Another aspect pertains to a host cell comprising a circularized double stranded DNA nucleic acid obtainable by the method. Further, therapeutic and non-therapeutic uses of a circularized double stranded DNA nucleic acid obtainable by the method. Finally, a kit for the circularization of a double stranded DNA nucleic acid.

A method of performing a non-isothermal nucleic acid amplification reaction, the method comprising the steps of: (a) mixing a target sequence with one or more complementary single stranded primers in conditions which permit a hybridization event in which the primers hybridize to the target, which hybridization event, directly or indirectly, leads to the formation of a duplex structure comprising two nicking sites disposed at or near opposite ends of the duplex; and performing an amplification process by; (b) using a nicking enzyme to cause a nick at each of said nicking sites in the strands of the duplex; (c) using a polymerase to extend the nicked strands so as to form newly synthesized nucleic acid, which extension with the polymerase recreates nicking sites; (d) repeating steps (b) and (c) as desired so as to cause the production of multiple copies of the newly synthesized nucleic acid.

The present disclosure provides improved nucleic acid sequencing-by-synthesis (SBS) methods, related kits and reagents, and systems for performing such methods using such kits and reagents.

The present invention relates to a detection method for detecting a target RNA contained in a sample with high sensitivity by using nicking/extension chain reaction system-based isothermal nucleic acid amplification (NESBA) that uses activity of a cleavage enzyme and a DNA polymerase. The NESBA of the present invention is a new concept isothermal target RNA detection method that realizes higher amplification efficiency than the existing NASBA technology and is deemed to be utilizable as a new concept diagnosis technology that can replace conventional target RNA detection technologies.

The present invention relates to a detection method for detecting a target RNA contained in a sample with high sensitivity by using nicking/extension chain reaction system-based isothermal nucleic acid amplification (NESBA) that uses activity of a cleavage enzyme and a DNA polymerase. The NESBA of the present invention is a new concept isothermal target RNA detection method that realizes higher amplification efficiency than the existing NASBA technology and is deemed to be utilizable as a new concept diagnosis technology that can replace conventional target RNA detection technologies.

Nucleic acid molecule analysis systems are described. The system may include a nucleic acid molecule attached to a particle with a characteristic dimension. The system may also include an aperture defined by a first electrode, a first insulator, and a second electrode. The aperture may have a characteristic dimension less than the characteristic dimension of the particle. The system may further include a first power supply in electrical communication with the first electrode and the second electrode. In addition, the system may include a second power supply configured to apply an electric field through the aperture. In some embodiments, the aperture may be defined by a first insulator. A portion of the first electrode may extend into the aperture. A portion of the second electrode may extend into the aperture.

Nucleic acid molecule analysis systems are described. The system may include a nucleic acid molecule attached to a particle with a characteristic dimension. The system may also include an aperture defined by a first electrode, a first insulator, and a second electrode. The aperture may have a characteristic dimension less than the characteristic dimension of the particle. The system may further include a first power supply in electrical communication with the first electrode and the second electrode. In addition, the system may include a second power supply configured to apply an electric field through the aperture. In some embodiments, the aperture may be defined by a first insulator. A portion of the first electrode may extend into the aperture. A portion of the second electrode may extend into the aperture.

Provided herein, among other things, is a conjugate comprising a polymerase and a nucleoside triphosphate, where the polymerase and the nucleoside triphosphate are covalently linked via a linker that comprises a cleavable linkage. A set of such conjugates, where the conjugates correspond to G, A, T (or U) and C is also provided. Methods for synthesizing a nucleic acid of a defined sequence are also provided. The conjugates can also be used for sequencing applications.

Provided herein, among other things, is a conjugate comprising a polymerase and a nucleoside triphosphate, where the polymerase and the nucleoside triphosphate are covalently linked via a linker that comprises a cleavable linkage. A set of such conjugates, where the conjugates correspond to G, A, T (or U) and C is also provided. Methods for synthesizing a nucleic acid of a defined sequence are also provided. The conjugates can also be used for sequencing applications.

A sequencing device is disclosed. The sequence device includes an array of conducting electrode pairs, each pair of electrodes comprising a source and a drain electrode arrangement separated by a nanogap, the electrode array deposited and patterned on a dielectric substrate; at least one transition metal dichalcogenide (TMD) layer disposed on each pair of electrodes, wherein the TMD layer connects each source and drain electrode within each pair, and bridges each nanogap of each pair of electrodes; and a dielectric masking layer disposed on the TMD layer and comprising at least one opening that defines an exposed TMD region, wherein the at least one opening is sized so as to allow a single biomolecule to fit therein and to attach on to the exposed TMD region. In embodiments of the disclosure, the TMD layer be a defective TMD layer.