Methods of detecting target polynucleotide sequences may include introducing one or more nucleic acid analytes to a first reaction mixture comprising a single-stranded probe oligonucleotide A.sub.0, a pyrophosphorolysing enzyme, and a ligase. The analyte may anneal to the single-stranded probe oligonucleotide A.sub.0 to create a first intermediate product which is at least partially double-stranded, where the 3′ end of A.sub.0 forms a double-stranded complex with the analyte and where A.sub.0 is pyrophosphorylsed in the 3′-5′ direction from the 3′ end to create at least a partially digested strand A.sub.1. A.sub.1 may undergo ligation to form oligonucleotide A.sub.2. The methods may also include detecting a signal derived from the formed oligonucleotides, and inferring therefrom the presence or absence of the target polynucleotide sequence in the analyte.

Methods and compositions are provided for the identification, detection, characterization, and/or utilization of double strand breaks in a target polynucleotide; the identification, detection, characterization, and/or utilization of cutting sites for double-strand-break-inducing agents; and the identification, detection, characterization, and/or utilization of double-strand-break-inducing agents.

Methods and compositions are provided for the identification, detection, characterization, and/or utilization of double strand breaks in a target polynucleotide; the identification, detection, characterization, and/or utilization of cutting sites for double-strand-break-inducing agents; and the identification, detection, characterization, and/or utilization of double-strand-break-inducing agents.

Provided are a vesicular adaptor and a single-chain cyclic library constructed by using the adaptor. The library can be used for RNA sequencing and other sequencing platforms dependent on a single-stranded cyclic library, and has the advantage of high throughput sequencing, high accuracy and simple operations.

Provided are a vesicular adaptor and a single-chain cyclic library constructed by using the adaptor. The library can be used for RNA sequencing and other sequencing platforms dependent on a single-stranded cyclic library, and has the advantage of high throughput sequencing, high accuracy and simple operations.

A method for rapidly preparing a Sanger sequencing template, comprising: performing denaturing treatment on a sample containing circular DNAs; performing rolling circle amplification of the denaturing product; and treating the rolling circle amplification product with alkaline phosphatase to remove residual dNTP.

A method for rapidly preparing a Sanger sequencing template, comprising: performing denaturing treatment on a sample containing circular DNAs; performing rolling circle amplification of the denaturing product; and treating the rolling circle amplification product with alkaline phosphatase to remove residual dNTP.

An apparatus suitable for single molecule sequencing. The apparatus includes at least one nanowell, a plurality of nucleic acid immobilization moieties, and a plurality of types of nucleic acid fragments. The nanowell has an observation zone. The nucleic acid immobilization moieties are disposed in or proximate to the observation zone. The nucleic acid fragments are immobilized to the nucleic acid immobilization moieties, respectively. At least one polymerase is disposed in the observation zone. A method of sequencing nucleic acid molecules using the above-mentioned apparatus is provided.

The invention relates to the use of polyphosphate containing species in a method of nucleic acid synthesis, to methods of nucleic acid synthesis, and to the use of kits comprising said polyphosphate containing species. The invention also relates to the use of polyphosphate containing species for the capping of 3-terminal hydroxyl moieties using terminal transferases.

The invention relates to the use of polyphosphate containing species in a method of nucleic acid synthesis, to methods of nucleic acid synthesis, and to the use of kits comprising said polyphosphate containing species. The invention also relates to the use of polyphosphate containing species for the capping of 3-terminal hydroxyl moieties using terminal transferases.