The present invention provides, among other things, methods for purifying mRNA based on normal flow filtration for therapeutic use.

The present disclosure provides a method of purifying mRNA molecules comprising (Ia) purifying precipitated mRNA molecules from a suspension comprising precipitated mRNA molecules, (Ib) washing and dissolving the purified precipitated mRNA molecules, (IIa) purifying the mRNA molecules using a solution comprising a chelating agent, followed by (IIb) washing the purified mRNA molecules, wherein steps (Ia) to (IIb) are performed using tangential flow filtration.

Methods for preparing microRNAs from microvesicles isolated from a biological sample from a subject, and preparation of DNA from microvesicle microRNA preparations.

Methods are provided for the stabilization and separation of nucleic acids from a sample via contact of the sample with a lysis and stabilization reagent that includes a cationic detergent. The cationic detergent lyses cells in the sample and stabilizes the released nucleic acids via the formation of nucleic acid-surfactant (NAS) complexes. The NAS complexes are centrifugally precipitated, washed, the resuspended in an aqueous resuspension liquid, forming a NAS complex suspension. The suspension is thermally processed to disintegrate the NAS complexes, thereby releasing the nucleic acids and forming a nucleic acid solution. In some example embodiments, the aqueous resuspension liquid is selected to be suitable for performing molecular amplification assays, such that the nucleic acid solution may be employed for performing a molecular amplification assay in the absence of further nucleic acid extraction. Examples are provided whereby the present methods are adapted for performing transcriptomic biomarker assays.

The present disclosure is directed to a method for purifying a sample containing nucleic acids to obtain isolated nucleic acids of a desired size range, either above a size cut-off, below a cut-off, or within a defined band of sizes, including: a) combining a nucleic acid-containing sample with a binding buffer to provide a binding mixture; b) contacting the binding mixture with a silica nanomembrane, wherein the silica nanomembrane adsorbs nucleic acids from the binding mixture within a desired size-range; and c) separating the bound nucleic acid from the remaining sample. Kits including a silica nanomembrane, a binding buffer and one or wash buffers are also provided herein.

In a nucleic acid extraction container, a filter part includes: a hydrophilic filter for collecting biological materials containing a nucleic acid from a sample; a biological material collection area for collecting biological materials on the filter; and a sample permeation area for allowing the passage of a sample having permeated through the filter. A sample injection port communicates with the biological material collection area. An air communication port communicates with the biological material collection area. A sample discharge port communicates with the sample permeation area. The air communication port is configured so as to be outwardly openable and closable.

In a nucleic acid extraction container, a filter part includes: a hydrophilic filter for collecting biological materials containing a nucleic acid from a sample; a biological material collection area for collecting biological materials on the filter; and a sample permeation area for allowing the passage of a sample having permeated through the filter. A sample injection port communicates with the biological material collection area. An air communication port communicates with the biological material collection area. A sample discharge port communicates with the sample permeation area. The air communication port is configured so as to be outwardly openable and closable.

The present invention provides, among other things, methods of purifying messenger RNA (mRNA) including the steps of (a) precipitating mRNA from an impure preparation; (b) subjecting the impure preparation comprising precipitated mRNA to a purification process involving membrane filtration such that the precipitated mRNA is captured by a membrane; and (c) eluting the captured precipitated mRNA from the membrane by re-solubilizing the mRNA, thereby resulting in a purified mRNA solution. In some embodiments, a purification process involving membrane filtration suitable for the present invention is tangential flow filtration.

Provided are methods and compositions for negatively and positively selecting for different size nucleic acid (e.g., DNA or RNA) fragments on borosilicate glass fiber membranes, silica and metal oxide surfaces such that only those fragments falling within a desired size range are obtained.

The present invention relates to an extraction apparatus, extraction method, and fluidic chip for extracting a target material. In the extraction apparatus according to the present invention, in a state in which a target material is bound to a porous film in an extraction kit, when an elution solution and a magnetic solution sequentially pass, the elution solution remaining in the porous film is pushed out by the magnetic solution due to a difference in polarity, and collected in a collection chamber. In addition, even in the collection chamber, the elution solution and the magnetic solution are maintained in a stacked state due to different polarities, and in a state in which the magnetic solution in the collection chamber is physically separated from the elution solution by magnetism of a magnetism applicator, the elution solution in the collection chamber may be recovered. Accordingly, after the elution solution is collected in the collection chamber along with the magnetic solution, in a state in which the magnetic solution in the collection chamber is physically separated from the elution solution by magnetism, the elution solution may be recovered.