The present invention is directed generally to the use of stem-loop oligonucleotides comprising an inverted repeat, a loop, and at least one degradable site or site capable of becoming a degradable site in the preparation of DNA molecules, such as a library. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.

A method for constructing a functional nucleic acid molecule comprising 1 or 2 nucleic acid strands, wherein 2 or more fragments having at corresponding ends a functional group pair that can mutually couple through a chemical reaction are introduced into a cell, and a functional nucleic acid molecule comprising 1 or 2 nucleic acid strands is formed by ligating mutually the fragments through a reaction between the functional groups in the cell.

Provided are a CAR library used to screen scFvs that can be functional in CAR-T cells, and an scFv manufacturing method in which the CAR library is used. A chimeric antigen receptor (CAR) library of the present invention includes nucleic acids coding for first CARs. Each of the first CARs includes a first antigen-binding domain, a first transmembrane domain, and a first intracellular signaling domain. The first antigen-binding domain includes a first single-chain antibody (scFv) to be screened for the ability to bind to a target antigen. The first scFv includes a first heavy-chain variable region and a first light-chain variable region. The first heavy-chain variable region and the first light-chain variable region meet a predetermined condition.

Disclosed herein are antigenic peptide-MHC molecules, termed comPACTs, and methods of producing such molecules. Also disclosed herein are methods of producing libraries of comPACT polynucleotides and polypeptides, and their exemplary use in capturing cancer neoepitope-reactive T cells.

This invention relates to the purification of nucleic acid conjugates, for example for use in DNA encoded chemical libraries. Reaction members that comprise nucleic acid conjugate reaction products and nucleic acid or nucleic acid conjugate reactants comprising a first reactive group are contacted with a capping molecule comprising a capture group. The capping molecule reacts with the first reactive group to form a covalent bond, thereby attaching the capture group to nucleic acid or nucleic acid conjugate reactants in reaction members. The nucleic acid or nucleic acid conjugate reactants are then removed from the reaction members using a binding member that binds the capture group, thereby producing a purified population of nucleic acid conjugate products. Methods of producing purified populations of nucleic acid conjugate products and nucleic acid chemical libraries are provided along with chemical libraries and kits.

A method of diagnosing severe sepsis prior to definitive clinical diagnosis. A pattern of gene expression that correlates strongly with a future diagnosis of severe sepsis and organ failure was identified in patients who had their blood drawn at first clinical presentation. The methods comprise identifying a pattern of two or more polynucleotides, whereby the altered expression of these polynucleotides correlates with prospective and actual sepsis. Also methods of identifying agents for treating sepsis based on the characteristics of this gene expression pattern are provided.

The invention provides for a diagnostic test to monitor cancer-specific genetic abnormalities to diagnose cervical cell disorders and predict which patients might progress to cancer. Genetic abnormalities are detected by identification in chromosomal copy number of chromosome 3 and 5 using FISH analysis of probes targeted to 3q and/or 5p.

Provided herein are methods of selective screening. In addition, various targeting proteins and sequences, as well as methods of their use, are also provided.

Provided herein are methods and compositions relating to CD3 libraries having nucleic acids encoding for a scaffold comprising a CD3 domain. CD3 libraries described herein encode for immunoglobulins such as antibodies.

Disclosed herein include methods, compositions, and kits suitable for use in generating libraries for nucleic acid sequencing. There are provided, in some embodiments, a plurality of protein complexes. Each protein complex can comprise a transposome and a programmable DNA binding unit capable of specifically binding to a user-selected binding site on a target double-stranded DNA (dsDNA). The binding site for each of the plurality of protein complexes can be different from each other. The transposome can comprise a transposase, a first adaptor, and a second adaptor. The first adaptor, the second adaptor, or both, can be a sequencing adaptor.