Disclosed herein, inter alia, are methods and compositions for sequencing a plurality of template nucleic acids.

Modifications to both hardware and enzymatic reactions used in single cell analyses such as but not limited to Seq-well that enable significant increases in the yield of transcripts per cell, portability and ease of use, increased scalability of the assay, and linkage of transcript information to other measurements made in the picowell arrays are disclosed.

This application provides a bead with a covalently attached chemical compound and a covalently attached DNA barcode and methods for using such beads. The bead has many substantially identical copies of the chemical compound and many substantially identical copies of the DNA barcode. The compound consists of one or more chemical monomers, where the DNA barcode takes the form of barcode modules, where each module corresponds to and allows identification of a corresponding chemical monomer. The nucleic acid barcode can have a concatenated structure or an orthogonal structure. Provided are method for sequencing the bead-bound nucleic acid barcode, for cleaving the compound from the bead, and for assessing biological activity of the released compound.

Provided are a rolling circle amplification method, a method for preparing a sequencing library, and a DNA nanoball prepared therefrom. The rolling circle amplification method includes: sequentially denaturing and annealing a double-stranded DNA and a mediating sequence in a same system, to complementarily pair the mediating sequence with two ends of a denatured single-stranded DNA; simultaneously introducing a ligase and a polymerase into the system to connect the two ends of the single-stranded DNA under action of the ligase; and performing a rolling circle amplification reaction under action of the polymerase by using the mediating sequence as a primer and the single-stranded DNA as a template, to obtain DNA nanoball.

The present disclosure relates to a biologics development platform that derives biologics from aptamers found to bind to a target. Particularly, aspects of the present disclosure are directed to generating sequencing data and analysis data for each unique aptamer of an aptamer library that binds to a target within a monoclonal compartment, inferring aptamer sequences derived from the sequencing data and the analysis data, identifying interaction points between the aptamer sequences and epitopes of the target based on structure or sequence motifs of the aptamer sequences, modeling molecular dynamics of interactions between the aptamer sequences and the epitopes to identify characteristics of the interaction points as requirements or restraints for the interactions, and inferring one or more amino acid sequences based on the characteristics of the interaction points derived from the interactions between aptamer sequences and the epitopes.

A nucleic acid patch method for amplifying target nucleic acid sequences in circulating free DNA or residual DNA samples where the defining ends of the target nucleic acid sequences are unknown.

Systems, methods, and compositions provided herein relate to preparation of beads encapsulating long DNA fragments for high-throughput spatial indexing. Some embodiments include preparation of nucleic acid libraries within the bead, wherein the bead includes pores that allow diffusion of reagents while retaining genetic material.

A synthesis and screening method of a DNA-encoded compound library. The DNA-encoded compound library consists of a DNA-encoded compound of formula (I). The screening method includes: incubating the DNA-encoded compound library with a protein target, followed by covalent cross-linking to obtain a covalently cross-linked complex; separating the covalently cross-linked complex from members in the library that do not bind to the protein target; and subjecting the covalently cross-linked complex to polymerase chain reaction (PCR) amplification and DNA sequencing.

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The present invention is directed to a method for the synthesis of a bi-functional complex comprising a molecule part and an identifier oligonucleotide part identifying the molecule part. A part of the synthesis method according to the present invention is preferably conducted in one or more organic solvents when a nascent bi-functional complex comprising an optionally protected tag or oligonucleotide identifier is linked to a solid support, and another part of the synthesis method is preferably conducted under conditions suitable for enzymatic addition of an oligonucleotide tag to a nascent bi-functional complex in solution.

The present disclosure provides targeted hybridization and/or proximity ligation of a probe for amplification and analysis of target sequences. The hybridization of the probe to the target sequences can be direct or through indirect association.