A CE based method and kit for the determination of the size and purity of an AAV genome which relies on Capillary Electrophoresis-Laser Induced Fluorescence (CE-LIF) analysis. These methods and kits are capable of detecting intact and partial genomes in a virus vectors such as adeno-associated viruses as well as remove small size impurities. In one example, the method can include creating a nucleic acid ladder using CE-LIF, releasing the genome from within an adeno-associated virus, purifying said genome and analyzing said genome using CE-LIF and comparing the results of the analysis of the genome to the nucleic acid ladder to determine a size of nucleic acids in the genome.

A CE based method and kit for the determination of the size and purity of an AAV genome which relies on Capillary Electrophoresis-Laser Induced Fluorescence (CE-LIF) analysis. These methods and kits are capable of detecting intact and partial genomes in a virus vectors such as adeno-associated viruses as well as remove small size impurities. In one example, the method can include creating a nucleic acid ladder using CE-LIF, releasing the genome from within an adeno-associated virus, purifying said genome and analyzing said genome using CE-LIF and comparing the results of the analysis of the genome to the nucleic acid ladder to determine a size of nucleic acids in the genome.

The invention provides compositions comprising nucleic acid complexes for use in monitoring binding interactions and in measuring association and/or dissociation kinetics, detecting analytes including low concentration analytes, and screening library members. In some instances, the nucleic acid complexes are double-stranded nicked nucleic acids comprising a scaffold nucleic acid hybridized to one or more oligonucleotides. In some instances, a first, a second, a third, and optionally a fourth oligonucleotide are linked to moieties that are known to interact with each other or which are suspected of interacting with each other or of interacting with a common moiety such as an analyte. Changes in topology of the complex are used to determine the binding interactions of the various binding partners.

The invention provides compositions comprising nucleic acid complexes for use in monitoring binding interactions and in measuring association and/or dissociation kinetics, detecting analytes including low concentration analytes, and screening library members. In some instances, the nucleic acid complexes are double-stranded nicked nucleic acids comprising a scaffold nucleic acid hybridized to one or more oligonucleotides. In some instances, a first, a second, a third, and optionally a fourth oligonucleotide are linked to moieties that are known to interact with each other or which are suspected of interacting with each other or of interacting with a common moiety such as an analyte. Changes in topology of the complex are used to determine the binding interactions of the various binding partners.

A nucleic acid (NA) detection method combines ultra-specific probe, on-chip isotachophoresis (ITP) which can separate single strand and double strand NAs, and enzyme amplification. The ITP device has a sieving matrix between the LE (leading electrolyte) and TE (trailing electrolyte) reservoirs, for separating double-strand and single-strand NAs. The LE or TE reservoir also contains a spacer ion having a mobility between the LE and the TE. The sample and a double-strand NA probe is added to the TE reservoir, the probe being formed of a protector strand modified with a fluorescent molecule and a complement strand, where the protector strand is released in the presence of the target NA. Fluorescent signal is detected downstream of the sieving matrix. Alternatively, the protector strand is modified with an enzyme and a single-strand NA modified with a substrate of the enzyme is added to the reaction mixture downstream of the sieving matrix.

A nucleic acid (NA) detection method combines ultra-specific probe, on-chip isotachophoresis (ITP) which can separate single strand and double strand NAs, and enzyme amplification. The ITP device has a sieving matrix between the LE (leading electrolyte) and TE (trailing electrolyte) reservoirs, for separating double-strand and single-strand NAs. The LE or TE reservoir also contains a spacer ion having a mobility between the LE and the TE. The sample and a double-strand NA probe is added to the TE reservoir, the probe being formed of a protector strand modified with a fluorescent molecule and a complement strand, where the protector strand is released in the presence of the target NA. Fluorescent signal is detected downstream of the sieving matrix. Alternatively, the protector strand is modified with an enzyme and a single-strand NA modified with a substrate of the enzyme is added to the reaction mixture downstream of the sieving matrix.

The present invention relates to a method of visualizing biomolecules, having the steps of: a) providing a sample of immobilized biomolecules in a matrix and carry on the electrophoresis process; b) incubating the matrix of step a) in a solution containing a cyanine-derived molecule, for a time of 5 to 60 minutes, at room temperature, in a container preventing exposure to light, shaking the container at less of 75 rpm; c) transferring the matrix from step b) to a container with a solution having: at least one tetrazolium salt and incubating for a time of 15 to 120 minutes at room temperature under light exposure; d) removing the matrix with immobilized biomolecules from the previous step and washing with distilled water; and e) visualizing directly by the naked eye the biomolecules immobilized in the matrix.

The present invention relates to a method of visualizing biomolecules, having the steps of: a) providing a sample of immobilized biomolecules in a matrix and carry on the electrophoresis process; b) incubating the matrix of step a) in a solution containing a cyanine-derived molecule, for a time of 5 to 60 minutes, at room temperature, in a container preventing exposure to light, shaking the container at less of 75 rpm; c) transferring the matrix from step b) to a container with a solution having: at least one tetrazolium salt and incubating for a time of 15 to 120 minutes at room temperature under light exposure; d) removing the matrix with immobilized biomolecules from the previous step and washing with distilled water; and e) visualizing directly by the naked eye the biomolecules immobilized in the matrix.

A method in which a mutant gene present in a gene pool mixedly with a large number of wild-type genes can be simply, inexpensively and sensitively detected is developed and provided. A clamping probe that is connected to a target nucleic acid molecule in two regions of first and second target nucleic acid complementary regions so that a wild-type target nucleic acid molecule and a mutant-type target nucleic acid molecule can be distinguished from each other depending on a difference in complementarity to these target nucleic acid molecules is provided.

A method in which a mutant gene present in a gene pool mixedly with a large number of wild-type genes can be simply, inexpensively and sensitively detected is developed and provided. A clamping probe that is connected to a target nucleic acid molecule in two regions of first and second target nucleic acid complementary regions so that a wild-type target nucleic acid molecule and a mutant-type target nucleic acid molecule can be distinguished from each other depending on a difference in complementarity to these target nucleic acid molecules is provided.