This invention pertains to compositions and methods for detecting single and/or multiple nucleotide mismatches on DNA fragments in vitro by enzymatic cleavage using a mixture of mismatch endonucleases. Additionally, this invention pertains to the ability to detect successful genome editing events by programmable nucleases, e.g., TALENS, RGENs, or ZFNs.

This invention pertains to compositions and methods for detecting single and/or multiple nucleotide mismatches on DNA fragments in vitro by enzymatic cleavage using a mixture of mismatch endonucleases. Additionally, this invention pertains to the ability to detect successful genome editing events by programmable nucleases, e.g., TALENS, RGENs, or ZFNs.

A method for quantifying individual mature tRNA species, comprising: incubating mature tRNA in a buffer to remove the amino acids from the 3 end; annealing a DNA/RNA stem-loop adapter; ligating the annealed hybrid stem-loop adapter to the mature tRNA; and amplifying and quantifying the ligation product by TaqMan qRT-PCR.

A method for quantifying individual mature tRNA species, comprising: incubating mature tRNA in a buffer to remove the amino acids from the 3 end; annealing a DNA/RNA stem-loop adapter; ligating the annealed hybrid stem-loop adapter to the mature tRNA; and amplifying and quantifying the ligation product by TaqMan qRT-PCR.

[Problem] An objective of the present invention is to develop a new type of method for nucleic acid detection that can be suitably used for detection of short nucleic acids.

[Solution] The present invention provides a method for nucleic acid detection including i) hybridizing a detection target nucleic acid 1 and a primer for nucleic acid detection 3 to form a DNA-RNA complex, ii) removing a first polynucleotide segment 301 from the primer for nucleic acid detection 3 by degrading at least a portion of the DNA-RNA complex with a nuclease 4, and iii) hybridizing the primer for nucleic acid detection 3 to a template nucleic acid 2 to perform a reaction for amplifying the template nucleic acid 2 by a polymerase 5.

[Problem] An objective of the present invention is to develop a new type of method for nucleic acid detection that can be suitably used for detection of short nucleic acids.

[Solution] The present invention provides a method for nucleic acid detection including i) hybridizing a detection target nucleic acid 1 and a primer for nucleic acid detection 3 to form a DNA-RNA complex, ii) removing a first polynucleotide segment 301 from the primer for nucleic acid detection 3 by degrading at least a portion of the DNA-RNA complex with a nuclease 4, and iii) hybridizing the primer for nucleic acid detection 3 to a template nucleic acid 2 to perform a reaction for amplifying the template nucleic acid 2 by a polymerase 5.

Methods, systems, kits and compositions are described for quality control and quantitation of nucleic acid libraries of double stranded nucleic acid libraries prior to massively parallel sequencing. Electrophoretic separation within a channel using a detectably labeled single stranded sizing ladder may be used to define the molecular weight range and amount of the double stranded nucleic acids.

Methods, systems, kits and compositions are described for quality control and quantitation of nucleic acid libraries of double stranded nucleic acid libraries prior to massively parallel sequencing. Electrophoretic separation within a channel using a detectably labeled single stranded sizing ladder may be used to define the molecular weight range and amount of the double stranded nucleic acids.

A method of using an exchange-induced remnant magnetization (EXIRM) technique for label free detection of short strands of nucleotides and cancer biomarkers, such as DNA and microRNA strands, DNA/RNA-binding biomarkers, and cancer-specific antigens, with high sensitivity, high specificity, and broad dynamic range. The method may provide a label-free approach aimed to facilitate high reliability, and to require a minimum amount of biochemical reagents.

A method of using an exchange-induced remnant magnetization (EXIRM) technique for label free detection of short strands of nucleotides and cancer biomarkers, such as DNA and microRNA strands, DNA/RNA-binding biomarkers, and cancer-specific antigens, with high sensitivity, high specificity, and broad dynamic range. The method may provide a label-free approach aimed to facilitate high reliability, and to require a minimum amount of biochemical reagents.