Patent classifications
C12N15/10
RAPID PRECIPITATION-DRIVEN KILOBASE SIZE SELECTION OF HMW DNA
Provided herein are methods of purifying a sample containing nucleic acids to obtain isolated nucleic acids of a desired size range and methods of sequencing nucleic acids of a desired size range. The methods include a) combining a nucleic acid-containing sample with a precipitation buffer in a container to provide a precipitation mixture in which the precipitation buffer comprises water, a buffer, a salt, and polyvinyl pyrrolidinone (PVP) and/or Ficoll. The methods also include precipitating the nucleic acids in the precipitation mixture to provide a precipitated nucleic acid portion and a remaining sample portion. The precipitated nucleic acid portion predominantly comprises nucleic acid molecules above a selected size cutoff value and the remaining sample portion predominantly comprises nucleic acid molecules below the selected size cutoff value. The methods also include separating the precipitated nucleic acid portion from the remaining sample portion. Related compositions and kits are also provided herein.
Sample Preparation Device and Methods for Using Same
Aspects of the present disclosure include sample preparation cartridges including a cylindrical structure and one or more covers. The cylindrical structure further includes a top, a bottom, an annular wall, a plurality of cavities in the annular wall that form a plurality of open-sided chambers on the annular wall and one or more interconnections providing fluidic communication between the plurality of chambers. The one or more covers cover the open side of the plurality of chambers. Also provided is a cylinder housing comprising one or more magnets. The sample preparation cartridge is removably disposed into the cylinder housing or adjacent to the cylinder housing. Methods of using the sample preparation device are also provided.
COMPOSITIONS, METHODS, AND SYSTEMS FOR GENOME EDITING TECHNOLOGY
The present disclosure provides compositions, methods, and systems related to genome editing technology. In particular, the present disclosure provides a novel CRISPR-based genome editing technology that involves the generation of abasic sites to facilitate genetic recombination, without the need for breaks in the DNA. The compositions, methods, and systems described herein address many of the drawbacks of currently available approaches, including off-target effects and cellular toxicity.
HIGH-THROUGHPUT AUTOMATED STRAIN LIBRARY GENERATOR
A method for automated, high throughput cellular library generation is disclosed. The method includes providing a suspension including transformed cells and plating the transformed cells onto solid surfaces of each of at least one reservoir of a reservoir plate. The solid surfaces can include a liquid growth medium. The reservoir plate is incubated, and after cellular growth has occurred on at least one plated surface of the reservoir plate, a series of automatic steps are performed. The automatically-performed steps include adding disaggregation solution to the reservoir plate, applying a mechanical force, such as a rotational force, to the reservoir plate to produce resuspended cells, and/or collecting the resuspended cells.
In-vivo Continuous Directed Evolution System and Application Thereof
The disclosure discloses an in-vivo continuous directed evolution system and application thereof, and belongs to the fields of gene engineering and enzyme engineering. The system includes Escherichia coli host bacteria carrying a random mutation module mutagenesis plasmid, a programmed death module toxin-antitoxin system and a target gene expression module target plasmid. The modules are coupled with one another, and target genes are subjected to multiple rounds of continuous mutation by virtue of the random mutation module mutagenesis plasmid in the system, so that the mutation rate of the target genes is further increased, and ultimately, efficient evolution and screening of the target genes in the host bacteria are realized. According to the system, mutations are accurately positioned on the target genes, random mutations in non-target gene regions are reduced, and the system has good practical value and can be applied to directed evolution of various different functional proteins.
Target enrichment
The present disclosure provides, among other things, a way to amplify and sequence target sequences in a low-input sample. In some embodiments, the method comprises ligating a double-stranded adaptor onto a population of fragments to produce tagged fragments, and linearly amplifying the tagged fragments.
Methods for identifying selective binding pairs
The present disclosure methods for identifying binding partners using cell surface display libraries, where the cells of the library display engineered peptides on their cell surfaces for identification of peptides that bind to targets of interest. The engineered peptides are preferably expressed in the cells under conditions that provide both secretion and display of the engineered peptides on the cell surfaces, thus providing access of the engineered peptides to identify potential binding pairs. The cell libraries cab be engineered using an automated editing system that provides for one or more targeted edits per cell.
Water-soluble prolamin, preparation thereof, and uses thereof for food-grade application including prolamin compositions combining prolamin and DNA taggants
A water-soluble prolamin composition, such as a zein composition, and methods for producing the same. A method for tagging items comprising applying a plurality of non-coding DNA tags in a prolamin composition, such as a zein composition, wherein the selection of the particular taggants corresponds with a binary or nonbinary code sequence containing information about the tagged items.
METHOD FOR ENRICHING VESICULAR RNA
The present invention pertains to methods and kits for enriching extracellular nucleic acids such as vesicular RNA from a sample comprising extracellular vesicles. Accordingly to the methods an acidic binding mixture is prepared comprising the sample and anion exchange particles and binding extracellular vesicles to the anion exchange particles. After separating the anion exchange particles comprising the bound extracellular vesicles from the remaining mixture, bound extracellular vesicles are lysing the in the presence of at least one detergent and released RNA is bound to the anion exchange particles. The anion exchange particles with the bound RNA from the lysate are then eluted.
METHOD FOR PROCESSING POLYNUCLEOTIDE-CONTAINING SAMPLES
Methods and systems for processing polynucleotides (e.g., DNA) are disclosed. A processing region includes one or more surfaces (e.g., particle surfaces) modified with ligands that retain polynucleotides under a first set of conditions (e.g., temperature and pH) and release the polynucleotides under a second set of conditions (e.g., higher temperature and/or more basic pH). The processing region can be used to, for example, concentrate polynucleotides of a sample and/or separate inhibitors of amplification reactions from the polynucleotides. Microfluidic devices with a processing region are disclosed.