Provided are methods, as well as compositions, kits, and systems for preparing optimized control nucleic acids (polynucleotides) having reduce nucleic acid damage. Provided nucleic acid compositions provide reduced artifacts as compared to nucleic acid compositions prepared by conventional methods. Provided compositions are useful control in a variety of applications, including, but not limited to sequencing workflows to effectively monitor sensitivity, accuracy and/or precision of data.

The subject matter disclosed herein is generally directed to isolating single cells and nuclei from tissue samples for use in the analysis of single cells from archived biological samples. The subject matter disclosed herein is directed to isolating single cells and nuclei from formalin-fixed paraffin-embedded (FFPE) tissues. The subject matter disclosed herein is also directed to isolating single nuclei that preserve ribosomes or ribosomes and rough ER from frozen tissues. The subject matter disclosed herein is also directed to therapeutic targets, diagnostic targets and methods of screening for modulating agents.

The subject matter disclosed herein is generally directed to isolating single cells and nuclei from tissue samples for use in the analysis of single cells from archived biological samples. The subject matter disclosed herein is directed to isolating single cells and nuclei from formalin-fixed paraffin-embedded (FFPE) tissues. The subject matter disclosed herein is also directed to isolating single nuclei that preserve ribosomes or ribosomes and rough ER from frozen tissues. The subject matter disclosed herein is also directed to therapeutic targets, diagnostic targets and methods of screening for modulating agents.

The invention relates to an in vitro method for isolating nucleic acids associated to or contained inside extracellular vesicles (EVs) from a sample based on the formation of a DMB-EVs precipitate and the isolation of the nucleic acids present in the precipitate. The invention also relates to the use of the method of the invention for diagnosing or for determining the susceptibility of a subject to a disease, for determining the prognosis or for monitoring the progression of a disease, for monitoring the effect of a therapy, for identifying compounds suitable for the treatment of a disease, or for designing a personalized therapy or selecting a patient susceptible to being treated with a therapy for the prevention and/or treatment of a disease. In addition, the invention also relates to a kit comprising dimethylmethylene blue (DMB) and a reagent capable of isolating nucleic acids from EVs, and to its use.

The invention relates to an in vitro method for isolating nucleic acids associated to or contained inside extracellular vesicles (EVs) from a sample based on the formation of a DMB-EVs precipitate and the isolation of the nucleic acids present in the precipitate. The invention also relates to the use of the method of the invention for diagnosing or for determining the susceptibility of a subject to a disease, for determining the prognosis or for monitoring the progression of a disease, for monitoring the effect of a therapy, for identifying compounds suitable for the treatment of a disease, or for designing a personalized therapy or selecting a patient susceptible to being treated with a therapy for the prevention and/or treatment of a disease. In addition, the invention also relates to a kit comprising dimethylmethylene blue (DMB) and a reagent capable of isolating nucleic acids from EVs, and to its use.

Phosphoramidate-based monomers are provided for use in the synthesis of expandable polymers for nanopore-based sensing. Such monomers comprising a reporter construct that contain a first reporter code, a symmetrical chemical brancher bearing a translocation control element, and a second reporter code, wherein the ends of the reporter construct are attached to phosphoramidate-nucleoside. Related methods and products are also provided.

Phosphoramidate-based monomers are provided for use in the synthesis of expandable polymers for nanopore-based sensing. Such monomers comprising a reporter construct that contain a first reporter code, a symmetrical chemical brancher bearing a translocation control element, and a second reporter code, wherein the ends of the reporter construct are attached to phosphoramidate-nucleoside. Related methods and products are also provided.

An example of a sequencing kit includes a flow cell, an encapsulation matrix precursor composition, and a radical initiator. The flow cell includes a plurality of chambers and primers attached within each of the plurality of chambers. The encapsulation matrix precursor composition consists of a fluid, a monomer or polymer including a radical generating and chain elongating functional group, a radical source, and a crosslinker. The radical initiator is part of the encapsulation matrix precursor composition or is a separate component.

An example of a sequencing kit includes a flow cell, an encapsulation matrix precursor composition, and a radical initiator. The flow cell includes a plurality of chambers and primers attached within each of the plurality of chambers. The encapsulation matrix precursor composition consists of a fluid, a monomer or polymer including a radical generating and chain elongating functional group, a radical source, and a crosslinker. The radical initiator is part of the encapsulation matrix precursor composition or is a separate component.

The present invention is directed to compositions, methods and kits useful for the generation of nucleic acids from RNA templated and further nucleic acid amplification and detection. Specifically, the invention is directed to the generation and amplification of nucleic acids by reverse transcriptase polymerase chain reaction (RT-PCR). Provided are compositions and methods for improved amplification of nucleic acid molecules in a two-step, addition only RT-PCR procedure. The invention thus facilitates the rapid and efficient amplification of nucleic acid molecules and the detection and quantitation of RNA molecules, and is useful for a variety of research, industrial, medical and forensic purposes.