Disclosed is a method for determining a nucleotide sequence of a target nucleic acid. The method comprises: providing a pool of amplicons; sequencing each amplicon in the pool of amplicons to obtain sequence information of each amplicon; comparing a part of the sequence information of each amplicon with at least a part of the sequence of the target specific primer section, wherein the part of the sequence information of each amplicon is a sequence starting from position X-y and y is positive integer; determining whether the part of the sequence information of each amplicon comprises at least the part of the sequence of the target specific primer section; and determining accurate sequence of the target region using sequence information which comprises at least the part of the sequence of the target-specific primer section.

The present invention relates to a method of detecting a target nucleic acid on the basis of rolling circle amplification (RCA), and more specifically, to a method of detecting a target nucleic acid, the method in which a target nucleic acid (a nucleic acid having a target nucleic acid sequence), when present, forms a circular template with a template for performing an amplification reaction, wherein during the amplification reaction, a restriction enzyme is added to further induce a new RCA reaction, thus increasing the reaction rate and sensitivity, and to an RCA composition for implementing the method. The method of detecting a target nucleic acid according to the present invention, by detecting a barcode sequence predefined according to the type of the target nucleic acid, enables multiple detections of the presence of the target nucleic acid without sequencing, is inexpensive for not using costly enzymes, such as CRISPR, can detect barcode sequences, and can utilize various existing nucleic acid detection systems, and thus, can be useful in the detection of gene mutations.

The present invention relates to a method of detecting a target nucleic acid on the basis of rolling circle amplification (RCA), and more specifically, to a method of detecting a target nucleic acid, the method in which a target nucleic acid (a nucleic acid having a target nucleic acid sequence), when present, forms a circular template with a template for performing an amplification reaction, wherein during the amplification reaction, a restriction enzyme is added to further induce a new RCA reaction, thus increasing the reaction rate and sensitivity, and to an RCA composition for implementing the method. The method of detecting a target nucleic acid according to the present invention, by detecting a barcode sequence predefined according to the type of the target nucleic acid, enables multiple detections of the presence of the target nucleic acid without sequencing, is inexpensive for not using costly enzymes, such as CRISPR, can detect barcode sequences, and can utilize various existing nucleic acid detection systems, and thus, can be useful in the detection of gene mutations.

A method for determining sequences from sense and antisense strands of a nucleic acid, including (a) providing a nucleic acid cluster attached to a solid support, wherein the nucleic acid cluster includes a sense strand and an antisense strand of a concatemer, the concatemer including multiple copies of a sequence unit, the sequence unit including a target sequence and a primer binding site; (b) hybridizing a primer to a primer binding site in the antisense strand; (c) extending the primer along the antisense strand to determine the sequence from at least a portion of the target sequence in the antisense strand; (d) hybridizing a second primer to a primer binding site in the sense strand; and (e) extending the second primer along the sense strand to determine the sequence from at least a portion of the target sequence in the sense strand.

A method for determining sequences from sense and antisense strands of a nucleic acid, including (a) providing a nucleic acid cluster attached to a solid support, wherein the nucleic acid cluster includes a sense strand and an antisense strand of a concatemer, the concatemer including multiple copies of a sequence unit, the sequence unit including a target sequence and a primer binding site; (b) hybridizing a primer to a primer binding site in the antisense strand; (c) extending the primer along the antisense strand to determine the sequence from at least a portion of the target sequence in the antisense strand; (d) hybridizing a second primer to a primer binding site in the sense strand; and (e) extending the second primer along the sense strand to determine the sequence from at least a portion of the target sequence in the sense strand.

The invention provides methods, compositions, kits and devices for the detection of target molecules. In some embodiments, the invention allows for multiplexed target molecule detection.

The invention provides methods, compositions, kits and devices for the detection of target molecules. In some embodiments, the invention allows for multiplexed target molecule detection.

The present disclosure provides a kit for preparing a library of nucleic acids. The kit includes first and second oligonucleotide, each having a tail sequence, a common sequence, and at least one of a unique identifier sequence, and a variable length punctuation mark. The kit further includes a first primer having a first sample identifier sequence and a first priming sequence at a 3′ end of the first primer. The first priming sequence includes the tail sequence of the first oligonucleotide. The kit further includes a second primer having a second sample identifier sequence and a second priming sequence at a 3′ end of the second primer. The second priming sequence is complimentary to the second tail sequence of the second oligonucleotide.

The present disclosure provides a kit for preparing a library of nucleic acids. The kit includes first and second oligonucleotide, each having a tail sequence, a common sequence, and at least one of a unique identifier sequence, and a variable length punctuation mark. The kit further includes a first primer having a first sample identifier sequence and a first priming sequence at a 3′ end of the first primer. The first priming sequence includes the tail sequence of the first oligonucleotide. The kit further includes a second primer having a second sample identifier sequence and a second priming sequence at a 3′ end of the second primer. The second priming sequence is complimentary to the second tail sequence of the second oligonucleotide.

Methods of producing an amplified double stranded deoxyribonucleic acid (dsDNA) from a nucleic acid sample are provided. Aspects of the methods include amplifying using a single product nucleic acid primer and a template switch oligonucleotide to produce an amplified dsDNA product. Compositions and kits for use in performing the methods are also provided.