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 research, diagnostics, and treatment.

A method is provided for enriching extracellular DNA from a biological sample comprising extracellular DNA and extracellular vesicles, wherein the method comprises: (a) preparing a binding mixture comprising—the biological sample, —a solid phase comprising anion exchange groups, —an acidic binding buffer comprising a buffering agent, and binding extracellular DNA to the solid phase comprising anion exchange groups; (b) separating the solid phase with the bound extracellular DNA from the remaining binding mixture, wherein the remaining binding mixture comprises extracellular vesicles. The method may furthermore comprise processing the remaining binding mixture to enrich one or more biological targets of interest therefrom, wherein processing may comprise (c) enriching as biological targets extracellular vesicles and/or extracellular RNA from the remaining binding mixture.

The present disclosure concerns the use of a mating pair stabilization module comprising a type IV adhesion pilus with a conjugative bacterial host cell or as a part of a conjugative delivery system for mediating effective in vivo conjugation.

A porous membrane obtainable by a process comprising curing a composition comprising: (i) cross-linking agent(s) comprising at least one ligand group; (ii) inert solvent(s); (iii) polymerization initiator(s); and (vi) optionally monomer(s) other than component (i) which are reactive with component (i); wherein the composition satisfies the following equation: Z=wt(i)/(wt(i)+wt(iii)+wt(iv)) wherein: Z has a value of at least 0.6; wt(i) is the number of grammes of component (i) present in the composition; wt(iii) is the number of grammes of component (iii) present in the composition; and wt(iv) is the number of grammes of component (iv) present in the composition.

Provided herein are methods for enhanced specificity of multiplexed measurements. Methods provided herein include immunoassay reactions and/or measuring protein-protein interactions with direct sequencing readouts of DNA barcodes.

A method for developing capture agents for target proteins employs a compound library to find cyclic peptide sequences that bind the target protein. The target protein is also reacted with a clickable group-provider reagent to provide the protein with clickable groups. The compounds in the library are provided with complementary clickable groups that bind the clickable group on the target protein when the peptide sequences bind the target protein. In some embodiments, the cyclic peptide sequences that bind the target protein are incorporated into constructs having one or more arms that can serve as capture agents or potential treatments against the pathogens from which the target protein is derived. Some embodiments provide pharmaceutical compositions for immunoassays, diagnostics, therapeutics or the like, that employ the constructs.

The present invention is directed to a method for extraction of cell-free nucleic acid fragments from a blood sample to facilitate cancer diagnosis, prognosis and monitoring as well as prenatal screening. The present invention provides a cartridge comprising a first compartment for filtering plasma from a blood sample and preferably also for cell fixation and cell rinsing in order to improve yield and a second compartment for performing nucleic acid separation, wherein the first compartment comprises a hollow fiber membrane and the second compartment comprises material for binding the nucleic acids or a gel for electrophoresis. The invention also provides and an automated system comprising a device with a docking site adapted to receive said cartridge, said device comprising means adapted to operate the blood plasma filtering process in said cartridge and means adapted to operate nucleic acid separation in said cartridge.

The present invention provides modified single primer extension-based methods for generating an amplified library of fragments of a target gene or genome of interest from a sample of fragmented DNA, wherein the library is suitable for use in detecting, quantifying and/or sequencing the target gene or genome of interest. The present invention also provides compositions for use in such methods. In some embodiments the present invention provides methods and compositions specifically for detecting, quantifying and/or sequencing circulating tumor derived HPV DNA.

The invention provides a method for synthesizing a DNA sequence comprising repeat units, including designing and synthesizing an extension primer and a blocking primer based on the repeat unit, performing a PCR amplification reaction by using the repeat unit (as an amplification template), the extension primer, and the blocking primer in a PCR reaction system, to obtain the DNA sequence comprising repeat units. The invention also provides a kit for this method. The method of the invention has the characteristics such as controllable copy number for repeat synthesis, simple synthesis steps, and low cost, and is very suitable for high-throughput production in industry.

Disclosed are a method and a device for constructing an antibody complementarity determining region (CDR) library. Also disclosed are a method, a device and a computer program product for determining the occurrence frequency of member sequences of an antibody CDR library, by means of which an antibody CDR library with a specific amino acid distribution at one or more positions can be obtained.