The present disclosure provides cell lysis buffers and method of use. Cell lysis buffers may comprise an aqueous solution comprising a buffering agent, a detergent, and cyanate or thiocyanate. The cell lysis buffers may be free or substantially free of urea and/or carbonic acid. Such lysis buffers may be used to lyse cells, e.g., in tissues. The cell lysis buffers may be suitable for use with isotachophoresis.

Disclosed are methods for purifying polynucleotides from a feed solution using oligonucleotide affinity columns at flowrates between 0.5 CV/min to 1000 CV/min. The methods can include several steps including loading a mixture containing the target polynucleotide onto a chromatography media, e.g., a column, that carries a macroporous support with an oligonucleotide affinity ligand bonded to a surface. The affinity ligand can hybridize the targeted polynucleotide and allow for separation and purification of the target.

Methods for preparing enriched sequencing libraries from test samples that contain double-stranded deoxyribonucleic acid (dsDNA) are provided.

Various aspects of the present disclosure are directed toward methods and apparatuses for interacting a first liquid and a second liquid in one or more fluidic channels of a capillary structure. The methods and apparatuses can include providing at least one capillary barrier that positions a meniscus of the first liquid at a fluid-interface region using capillary forces within the capillary structure. Additionally, a path is provided along one of the channels for the second liquid to flow toward the fluid-interface region. Additionally, gas pressure is released, via a gas-outflow port, from the fluid-interface region while flow of the first liquid is arrested. Further, the first liquid and the second liquid contact in the fluid-interface region with the capillary barrier holding the first liquid at the fluid-interface region.

Apparatus for processing a liquid comprising a target substance, preferably a biomolecule, is provided. The apparatus comprises at least a first and a second means for carrying out a unit operation, each means for carrying out a unit operation comprising a feed for a liquid in fluid connection with the inlet of a flow-controller comprising a variable flow inlet valve and an outlet, wherein at least one of the means for carrying out a unit operation comprises feeds for at least two liquids, the feeds being in fluid connection with the inlets of a multiple inlet flow-controller comprising two or more variable flow inlet valves for dosing the at least two liquids, the flow-controller also comprising an outlet; a feed for a liquid feedstock comprising the target substance in fluid connection with the outlet from the flow-controller thereby to enable combination of the feed for a liquid comprising the target substance with the mixed bioprocessing liquids to produce a device feed; a device for achieving a processing operation comprising a device inlet and a device outlet, the device inlet being in fluid connection with the device feed; and a means for imparting flow through the flow controller and from the feed for the liquid feedstock through the processing device via the device inlet; and wherein the feed for a liquid feedstock comprising the target substance for the second means for carrying out a unit operation comprises the outlet from the first means for carrying out a unit operation.

The present invention relates to a ribonucleic acid (RNA) extraction method using a solid subject, the method including activating a subject with a reactive amine group, injecting a sample having 110.sup.1 to 110.sup.3 cells/ml and a dimethyl suberimidate (DMS) compound or a dimethyl pimelimidate (DMP) compound into the subject, and forming a complex having the RNA within the sample and the compound, and extracting the RNA by treating elution buffer to the subject on which the complex is formed. The subject, particularly, the thin film device used for extracting the RNA, has improved hydrophilicity compared to the conventional silicon substrate, so that RNA is extracted more efficiently.

Ligands and methods for selectively binding hypermethylated DNA from a sample. The ligands include a CG-region binding molecule-conjugated resin derived from the aminoglycoside antibiotic amikacin. Furthermore, the CG-region binding molecule may be conjugated to the resin with a crosslinker and/or may be modified with one or more of long chain and short chain alkyl, aryl, piperazinyl, piperidyl, and pyrrolidyl groups. Such ligands are used in methods for contacting a sample to thereby selectively bind hypermethylated DNA.

The present invention relates to a method for detecting at least one oligonucleotide conjugate of interest in solution, wherein the oligonucleotide conjugate of interest is composed of a nucleic acid entity and of a nonpolar entity, wherein the nucleic acid entity is chemically linked to the nonpolar entity, and wherein the method comprises the steps of providing a liquid sample comprising the oligonucleotide conjugate of interest; separating the oligonucleotide conjugate of interest from the liquid sample by analytical means under conditions including the presence of at least one cyclodextrine in solution; and detecting the oligonucleotide conjugate of interest by means of qualitative or quantitative analysis.

A sample containing particles having high-molecular-weight (HMW) DNA is entrapped in a gel matrix, and the gel matrix is exposed to a lysis reagent configured to release the HMW DNA from the particles. The HMW DNA may be purified by subjecting the gel matrix to an electrophoretic field that removes the HMW DNA from the particles, lysis reagents, and/or other sample constituents, from the gel matrix such that the HMW DNA remains. The gel matrix may be subjected with DNA cleavase reagents configured to cleave at specific DNA sequences within the HMW DNA to liberate defined segments of the DNA as fragments of reduced size. The gel matrix may also be subjected to an electrophoretic field, which moves and separates the DNA fragments from uncleaved DNA of the HMW DNA, which remains substantially immobile. The electrophoretically separated DNA fragments may be isolated from the gel matrix.

Provided is a method for large-scale preparation of a purified preparation of a recombinant lentiviral vector at the GMP grade. The method comprises: (a) providing raw material feed liquid to be purified that comprises recombinant viral vectors; (b) carrying out a microfiltration treatment on the feed liquid to obtain a microfiltered filtrate comprising the recombinant viral vectors; (c) optionally concentrating the filtrate to obtain a concentrated filtrate; (d) purifying the filtrate obtained in the previous step by means of chromatography to obtain a crude pure product comprising the recombinant viral vectors; and (e) subjecting the crude pure product obtained in the previous step to liquid exchange and elaborate purification to obtain the purified recombinant viral vectors.