Methods, assays, compositions and kits for the ligation of short polynucleotides are presented herein. The short polynucleotides are optionally no more than 7 nucleotides in length, and can be as short as 3 or 4 nucleotides in length. The ligation is optionally performed by CV ligase.

Methods, assays, compositions and kits for the ligation of short polynucleotides are presented herein. The short polynucleotides are optionally no more than 7 nucleotides in length, and can be as short as 3 or 4 nucleotides in length. The ligation is optionally performed by CV ligase.

A target DNA sequence (1) is contacted with M.sub.L+M.sub.R ligation oligonucleotides (10, 20) under hybridization conditions. The ligation oligonucleotides (10, 20) comprises a respective UMI (14, 15). A ligating agent ligates together the ligation oligonucleotides (10, 20) while hybridized to the target DNA sequence (1) to form a ligated product (30). The ligated product (30) is amplified by means to amplification primers (40, 50), of which one comprises a sample-specific barcode sequence (55), to form an amplified product (60) comprising two UMIs (65), a sequence of interest (66) and one barcode sequence (68). Amplified products (60) from multiple samples are pooled together, sequenced, demultiplexed and mapped to enable quantification of unique target DNA sequences (1) in the different samples.

A method and kits are provided for nucleic acid quantification and discrimination using surface plasmon resonance (SPR). The method provided is able to significantly enhance the detection limit and multiplex the discrimination assay using the melting properties of the target DNA on top of standard PCR reaction. By using the heating and cooling cycles of the polymerase chain reaction (PCR) or Ligation chain reaction (LCR), DNA is melted and hybridized onto the SPR sensor surface together with a nanoparticle label. Thus, during every cycle of DNA amplification, the quantity and type of target DNA can be monitored.

A method and kits are provided for nucleic acid quantification and discrimination using surface plasmon resonance (SPR). The method provided is able to significantly enhance the detection limit and multiplex the discrimination assay using the melting properties of the target DNA on top of standard PCR reaction. By using the heating and cooling cycles of the polymerase chain reaction (PCR) or Ligation chain reaction (LCR), DNA is melted and hybridized onto the SPR sensor surface together with a nanoparticle label. Thus, during every cycle of DNA amplification, the quantity and type of target DNA can be monitored.

A nucleic acid probe and a nucleic acid sequencing method for performing sequencing while ligating nucleic acids. The nucleic acid probe is a DNA sequencing probe, comprising a first moiety, a second moiety, a linker, and a detectable label. A base of the first moiety is A, T, U, C, or G, a base of the second moiety is a random base and/or a universal base, and 3 bases or more are present in the second moiety. The first moiety and the second moiety are ligated via the linker, the connection between the first moiety and the ligation can be cleaved, and the detectable label is ligated to the second moiety or the linker. The above probe, a combination formed therewith, or a sequencing method using the same can reduce the number or types of probes in nucleic acid sequencing, thereby reducing cost.

A nucleic acid probe and a nucleic acid sequencing method for performing sequencing while ligating nucleic acids. The nucleic acid probe is a DNA sequencing probe, comprising a first moiety, a second moiety, a linker, and a detectable label. A base of the first moiety is A, T, U, C, or G, a base of the second moiety is a random base and/or a universal base, and 3 bases or more are present in the second moiety. The first moiety and the second moiety are ligated via the linker, the connection between the first moiety and the ligation can be cleaved, and the detectable label is ligated to the second moiety or the linker. The above probe, a combination formed therewith, or a sequencing method using the same can reduce the number or types of probes in nucleic acid sequencing, thereby reducing cost.

Disclosed are compositions, methods and kits for determining the presence, absence, amount, copy number, or other characteristics of one or more polynucleotide sequences in two or more samples and use thereof in genotyping, evaluation of copy number variation, expression analysis, determination of splice variants and fusion genes, and other genetic analyses.

Disclosed are compositions, methods and kits for determining the presence, absence, amount, copy number, or other characteristics of one or more polynucleotide sequences in two or more samples and use thereof in genotyping, evaluation of copy number variation, expression analysis, determination of splice variants and fusion genes, and other genetic analyses.

The invention relates to methods of detecting a genetic variation in a genetic sample from a subject using labeled probes and counting the number of labels in the probes.