A method includes (i) providing an RNA polynucleotide; (ii) modifying the RNA polynucleotide by annealing and ligating a polynucleotide comprising a 3′ terminal random multimer segment and having a stem-loop form; (iii) optionally performing a reverse transcription of the RNA polynucleotide; (iv) cleaving the stem-loop segment of the annealed polynucleotide to yield a 3′ A overhang; (v) connecting an adaptor polynucleotide complex associated with an RNA translocase enzyme and at least one cholesterol tether segment to the polynucleotide obtained in step (iv); (vi) contacting the modified RNA polynucleotide obtained in step (v) with a transmembrane pore such that the RNA translocase controls the movement of the RNA polynucleotide through the transmembrane pore and the cholesterol tether anchors the RNA polynucleotide in the vicinity of the transmembrane pore; and (vii) taking one or more measurements during the movement of the RNA polynucleotide through the transmembrane pore Other features are also disclosed.

A method includes (i) providing an RNA polynucleotide; (ii) modifying the RNA polynucleotide by annealing and ligating a polynucleotide comprising a 3′ terminal random multimer segment and having a stem-loop form; (iii) optionally performing a reverse transcription of the RNA polynucleotide; (iv) cleaving the stem-loop segment of the annealed polynucleotide to yield a 3′ A overhang; (v) connecting an adaptor polynucleotide complex associated with an RNA translocase enzyme and at least one cholesterol tether segment to the polynucleotide obtained in step (iv); (vi) contacting the modified RNA polynucleotide obtained in step (v) with a transmembrane pore such that the RNA translocase controls the movement of the RNA polynucleotide through the transmembrane pore and the cholesterol tether anchors the RNA polynucleotide in the vicinity of the transmembrane pore; and (vii) taking one or more measurements during the movement of the RNA polynucleotide through the transmembrane pore Other features are also disclosed.

The disclosure provides methods of synthesizing DNA using topoisomerase-mediated ligation, by adding single nucleotides or oligomers to a DNA strand in the 3′ to 5′ direction.

The disclosure provides methods of synthesizing DNA using topoisomerase-mediated ligation, by adding single nucleotides or oligomers to a DNA strand in the 3′ to 5′ direction.

Method of utilizing a nanochannel in combination with at least one magnetic sensor for detecting (e.g., identifying) molecules, cells, and other analytes. Particularly, the method includes bringing molecules, labeled with magnetic nanoparticles (MNPs), in close proximity to the magnetic sensor to identify the molecules via an output signal from the magnetic sensor. The method is particularly suited for identifying nucleotides of DNA and RNA strands.

Method of utilizing a nanochannel in combination with at least one magnetic sensor for detecting (e.g., identifying) molecules, cells, and other analytes. Particularly, the method includes bringing molecules, labeled with magnetic nanoparticles (MNPs), in close proximity to the magnetic sensor to identify the molecules via an output signal from the magnetic sensor. The method is particularly suited for identifying nucleotides of DNA and RNA strands.

In some embodiments, a computer-implemented method of determining an identity of one or more monomer subunit residues of a polymer analyte is provided, In some embodiments, a raw current signal generated by using a variable voltage to translocate the polymer analyte through a nanopore. In some embodiments, change points are detected in the raw current signal to determine a series of states, In some embodiments, capacitance compensation is performed on the raw current signal for each state to create an ionic current-vs-voltage curve for each state. In some embodiments, the ionic current-vs-voltage curves is converted to conductance-vs-voltage curves. In some embodiments, filtering is performed for the series of states to create a series of filtered states. In some embodiments, the identity of one or more monomer subunit residues of the polymer analyte is determined based on the series of filtered states.

In some embodiments, a computer-implemented method of determining an identity of one or more monomer subunit residues of a polymer analyte is provided, In some embodiments, a raw current signal generated by using a variable voltage to translocate the polymer analyte through a nanopore. In some embodiments, change points are detected in the raw current signal to determine a series of states, In some embodiments, capacitance compensation is performed on the raw current signal for each state to create an ionic current-vs-voltage curve for each state. In some embodiments, the ionic current-vs-voltage curves is converted to conductance-vs-voltage curves. In some embodiments, filtering is performed for the series of states to create a series of filtered states. In some embodiments, the identity of one or more monomer subunit residues of the polymer analyte is determined based on the series of filtered states.

An analyzing apparatus for molecules is provided. A pore device includes a cation selective nanopore, and a first chamber and a second chamber which are separated by the cation selective nanopore. In an initial state, the first chamber includes molecules to be analyzed, and the second chamber has higher salt concentration than the first chamber. A current sensor measures an ionic current flowing through a first electrode and a second electrode provided in the first chamber and the second chamber.

Methods of detecting a target nucleic acid sequence analyte are provided in which a crRNA and Cas12 or Cas13 enzyme are contacted to form a non-activated RNP. The non-activated RNP is contacted with a sample containing or suspected of containing the target nucleic acid sequence, and the target nucleic acid sequence and non-activated RNP specifically bind to each other if the target nucleic acid is present in the sample, thereby forming an activated RNP. A reporter nucleic acid is contacted with the activated RNP, and the activated RNP indiscriminately cleaves the reporter nucleic acid, reducing passage of intact, non-cleaved reporter nucleic acid through a nanopore in of a nanopore counting device such that a reduction of resistive pulses is produced which provides a signal representative of presence of the target nucleic acid sequence in the sample.