Provided herein is a method to reduce the endotoxin contamination in plasmid preparations. In the described method, plasmid DNA and endotoxins are bound to an anionic exchange matrix and are brought into contact with a wash buffer, comprising a combination of branched secondary alcohol ethoxylates with varying ethylene oxide chain lengths, wherein the branched secondary alcohol ethoxylates with the shorter ethoxylate chain is present in the washing buffer in excess compared to the branched secondary alcohol ethoxylate with the longer ethoxylate chain. The resulting purified plasmid has minimal endotoxin contamination levels and is considered endotoxin-free. Furthermore, provided are wash buffers comprising a combination of branched secondary alcohol ethoxylates with varying ethylene oxide chain lengths, kits comprising such wash buffers and the use of such wash buffers for reducing the endotoxin contamination in plasmid preparations.

An apparatus and method for extracting nucleic acids such as DNA molecules from biological samples uses solid-state magnetic manipulation. A fluid sample and magnetic beads are placed in a vessel. The vessel is placed in a housing with an array of electromagnets mounted therein. The electromagnets are energized sequentially or in groups to move the magnetic beads through the fluid sample in a variety of patterns. The apparatus disclosed herein may be used as a measurement device to measure bead number density and modify magnetic patterns in order to deliver consistent dosages in bead number.

The invention provides barcode libraries and methods of making and using them including obtaining a plurality of nucleic acid constructs in which each construct comprises a unique N-mer and a functional N-mer and segregating the constructs into a fluid compartments such that each compartment contains one or more copies of a unique construct. The invention further provides methods for digital PCR and for use of barcode libraries in digital PCR.

The disclosure provides amino acid sequence variants of Bacillus thuringiensis (Bt) toxins and methods of producing the same. Some aspects of this disclosure provide methods for generating Bt toxin variants by continuous directed evolution. Some aspects of this disclosure provide compositions and methods for pest control using the disclosed variant Bt toxins.

The present disclosure provides methods and kits for tracking nucleic acid target origin by barcode tagging of the targets when they break into smaller fragments. Nucleic acid targets are captured in vitro by clonally localized nucleic acid barcode templates on a solid support. Millions of nucleic acid targets can be processed simultaneously in a massively parallel fashion without additional partition. These captured targets are broken into small fragments, and a target specific barcode sequence is tagged on each fragment as an identification of their original target. These nucleic acid target tracking methods can be used for a variety of applications in both whole genome sequencing and targeted sequencing.

The present invention relates to recombinant DNA technology, in particular to methods for assembling two or more double stranded (ds) nucleic acid molecules with overlapping terminal sequences. In particular, the present invention relates to the use of a thermolabile DNA polymerase II derived 3′-5′ exonuclease isolated from Moritella viscoa and a thermolabile DNA polymerase I of marine origin in multi DNA assembly processes.

The present invention relates to the treatment of diseases relating to proteins of the human microbiome metasecretome and, thus, to microbiome interactions, especially microbiome-host interactions. In particular the present invention relates to a method for identification of secreted peptides and proteins of the human microbiome. The present invention also relates to methods for generating a database of human microbiome metasecretome protein sequences. Furthermore, the present invention relates to a method for preparing a protein of the human microbiome metasecretome as well as to the use of such proteins in medicine.

Provided herein are compositions and methods for accurate and scalable Primary Template-Directed Amplification (PTA) nucleic acid amplification and sequencing methods, and their applications for mutational analysis in research, diagnostics, and treatment. Further provided herein are multiomics methods for parallel analysis of DNA, RNA, and/or proteins from single cells. Provided herein are methods of multiomic single-cell analysis comprising: (a) isolating a single cell from a population of cells; (b) sequencing a cDNA library comprising polynucleotides amplified from mRNA transcripts from the single cell; and (c) sequencing a genome of the single cell.

Provided are high throughput methods for physically linking cDNA molecules derived from mRNA molecules expressed by the same cell, and libraries of linked cDNA molecules produced by the methods. The methods comprise reverse transcribing mRNA from a single cell in a first container to produce cDNA molecules, and linking the cDNA molecules in a second container. The methods unexpectedly produced libraries of cDNA molecules with an increase in the number of molecules that are correctly linked to other molecules derived from the same cell.

The invention provides highly effective and versatile CRISPR/Cas protein variants, compositions, methods and uses thereof in gene editing. More specifically, the invention relates to PAM-reduced or PAM-abolished Cas proteins and chimeras, complexes and conjugates thereof, genetic editing systems and to therapeutic and non-therapeutic methods and uses of the PAM-reduced or PAM-abolished Cas proteins.