The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

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.

Disclosed are devices, systems and methods for direct measurement of polymerase activity. In one example, a device includes at least a first electrode and a second electrode, the first and second electrode being separated by a gap; and a polymerase with two attachment sites, one for attaching to the first electrode and a second for attaching to the second electrode, wherein the two attachment sites are separated by a distance of at least about 1 nm and the distance does not significantly change with conformational changes of the polymerase.

Disclosed are devices, systems and methods for direct measurement of polymerase activity. In one example, a device includes at least a first electrode and a second electrode, the first and second electrode being separated by a gap; and a polymerase with two attachment sites, one for attaching to the first electrode and a second for attaching to the second electrode, wherein the two attachment sites are separated by a distance of at least about 1 nm and the distance does not significantly change with conformational changes of the polymerase.

The present invention relates to improvements in methods of high throughput nucleic acid sequencing, and in particular to improvements to methods of carrying out extension reactions during pairwise sequencing. The present invention relates to a method for carrying out a strand resynthesis extension reaction during pairwise sequencing, wherein said strand resynthesis extension reaction is carried out between a first sequencing read and a second sequencing read, and wherein said strand resynthesis extension reaction extends one or more immobilised primers to copy a first template strand to generate a second immobilised template strand; characterised in that the strand resynthesis extension reaction is carried out using a non-thermostable strand displacement polymerase at a temperature of less than 55° C., preferably at 38° C.

The present invention relates to improvements in methods of high throughput nucleic acid sequencing, and in particular to improvements to methods of carrying out extension reactions during pairwise sequencing. The present invention relates to a method for carrying out a strand resynthesis extension reaction during pairwise sequencing, wherein said strand resynthesis extension reaction is carried out between a first sequencing read and a second sequencing read, and wherein said strand resynthesis extension reaction extends one or more immobilised primers to copy a first template strand to generate a second immobilised template strand; characterised in that the strand resynthesis extension reaction is carried out using a non-thermostable strand displacement polymerase at a temperature of less than 55° C., preferably at 38° C.

Provided herein are methods of amplifying nucleic acids. In particular, methods are provided for amplifying circular RNA molecules. In certain embodiments, circular DNA molecules for amplification are generated from circular RNA molecules. Provided herein are methods for amplifying a nucleic acid. In certain embodiments, a method comprises priming a circular RNA template molecule with one or more DNA primers and extending the primers with a reverse transcriptase to generate a cDNA strand that is a copy of the circular RNA molecule. In certain embodiments, the cDNA strand generated is linear.

Provided herein are methods of amplifying nucleic acids. In particular, methods are provided for amplifying circular RNA molecules. In certain embodiments, circular DNA molecules for amplification are generated from circular RNA molecules. Provided herein are methods for amplifying a nucleic acid. In certain embodiments, a method comprises priming a circular RNA template molecule with one or more DNA primers and extending the primers with a reverse transcriptase to generate a cDNA strand that is a copy of the circular RNA molecule. In certain embodiments, the cDNA strand generated is linear.