Provided herein are methods, reagents, and kits for the isolation of sperm cells from other cell types and the purification of sperm-associated DNA from a complex sample. In particular, methods, reagents, and kits are provided for the separation of intact sperm cells from epithelial cells via binding of sperm cells to a cellulose material, and the extraction of sperm-associated DNA therefrom.

Methods, compositions and systems to amplify targets from RNA samples. The methods comprise designing target-specific primers, converting RNA into cDNA by reverse transcription, adding a universal primer by using template switching, and amplifying the targets by using multiplex PCR. The methods, compositions and systems described herein may further include, or include the use of, next generation sequencing (NGS) to analyze the sequences and various mutations of the amplified targets.

The present invention pertains to a method for isolating extracellular nucleic acids from a sample, wherein said sample is optionally stabilized, by binding the extracellular nucleic acids to a solid phase which carries anion exchange groups, comprising the following steps: a. binding the extracellular nucleic acids to the solid phase in a binding mixture having a first pH which allows binding the extracellular nucleic acids to the anion exchange groups of the solid phase; wherein the sample makes up at least 85% of the volume of the binding mixture; b. separating the solid phase with the bound extracellular nucleic acids; c. optionally washing the extracellular nucleic acids; d. optionally eluting extracellular nucleic acids from the solid phase. The method has the advantage that large sample volumes can be processed and that extracellular nucleic acids can be isolated rapidly with a high yield. The method is particularly suitable for automatable processes.

Aspects of the disclosure relate to liquid chromatography (e.g., HPLC) methods which enable high resolution separations of polynucleotides having hydrophobic portions (e.g., polyadenylated nucleic acids, such as mRNA) based upon the hydrophobic character of the molecules (e.g., polyA tail length). In some embodiments, the disclosure describes liquid chromatographic methods for separating a nucleic acid having a hydrophobic portion (e.g., a polyadenylated nucleic acid, such as an mRNA) from a complex mixture by a mobile phase system that comprises an ion pairing agent selected from Tris, inorganic cations (including e.g., Na, Li, K, ammonium, etc.), biological buffers (e.g., MOPS, HEPES, PIPES, etc.), and other charged or hydrophilic moieties, and lacks conventional ion pairing agents (e.g., Triethylammonium acetate, TEAA). Accordingly, in some embodiments methods described by the disclosure are useful for assessing the quality of pharmaceutical preparations comprising nucleic acids.

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.

Methods, systems, and devices for sampling/isolating material from cells. An exemplary system may comprise a chip including an electrode array of sampling electrodes arranged along a surface of the chip. A cell-receiving area may be located adjacent the surface of the chip. The system also may comprise a tag array of tags supported by the chip and aligned with the electrode array. Each tag of the tag array may include an identifier that is unique to the tag within the tag array. Each tag may be configured to bind nucleic acids, or a capturing agent distinct from the tag may be aligned with each sampling electrode of the electrode array to capture a protein or other analyte of interest. The system further may comprise a control circuit configured to apply an individually controllable voltage to each sampling electrode of the electrode array and measure an electrical property of the sampling electrode.

A method of DNA fragmentation is provided. The method comprising incubating a semi-solid biological sample which comprises the DNA with an auxiliary domain-directed nuclease having a binding affinity and selectivity to pre-defined sites in the DNA so as to yield a DNA fragment-of-interest, to thereby fragment the DNA.

Methods and apparatus providing for the isolation of an unknown mutation from a sample comprising wild type nucleic acids and mutated nucleic acids through the application of time-varying driving fields and periodically varying mobility-altering fields to the sample within in an affinity matrix.

Methods and apparatus providing for the isolation of an unknown mutation from a sample comprising wild type nucleic acids and mutated nucleic acids through the application of time-varying driving fields and periodically varying mobility-altering fields to the sample within in an affinity matrix.

Methods and devices for isolating or enriching target molecules from a sample are provided herein. In some embodiments, methods and devices further involve detection, analysis and/or sequencing of a target molecule.