The invention is directed to methods for synthesizing oligonucleotides direction on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3′-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3′ hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Disclosed are methods and systems for determining the three-dimensional structure of chromatin in eukaryotic cells. More specifically, disclosed are methods and systems for obtaining chromatin structural information by surface immobilization that includes tethering crosslinked protein:DNA complexes and/or ligated DNA complexes to media such as beads, gels, and or matrices during the conformation capture assay. In general, the method includes flash freezing a cell such that the structural organization of the chromatin or other protein:DNA complexes is preserved, cryomilling the cell, producing cross-linked protein:DNA complexes by cutting the chromatin using a chemical, physical or enzymatic method, substantially immobilizing the cross-linked protein:DNA complexes, ligating the cross-linked protein:DNA complexes intramolecularly such that the ligated protein:DNA complexes represent structural organization of the chromatin; characterizing the ligated DNA by sequencing or other methods; and identifying any structural organization of the chromatin. The structural organization preferably includes information relating to interacting loci of the chromatin.

Provided herein are compositions, methods and systems relating to libraries of polynucleotides such that the libraries allow for accurate and efficient hybridization after binding to target sequences. Further provided herein are probes, blockers, additives, buffers, and methods that result in improved hybridization. Such compositions and methods are useful for improvement of Next Generation Sequencing applications, such as reducing off-target binding or reducing workflow times.

A continuous flow reactor, a method of performing a continuous flow reaction, and a method of controlling a moveable wall of a reaction chamber of a continuous flow reactor. The reactor comprising: an inlet; an outlet; and a reaction chamber, between the inlet and the outlet and providing a flow path therebetween, the reaction chamber having a moveable wall; the reactor further comprising: a pressure sensor configured to monitor a fluid pressure in the continuous flow reactor; and a controller, operable to adjust the position of the moveable wall, and thereby change a volume of the reaction chamber, based on the monitored fluid pressure.

Biomolecule arrays on a substrate are described which contain a plurality of biomolecules, such as coding nucleic acids and/or isolated polypeptides, at a plurality of discrete, isolated, locations. The arrays can be used, for example, in high throughput genomics and proteomics for specific uses including, but not limited molecular diagnostics for early detection, diagnosis, treatment, prognosis, monitoring clinical response, and protein crystallography.

Biomolecule arrays on a substrate are described which contain a plurality of biomolecules, such as coding nucleic acids and/or isolated polypeptides, at a plurality of discrete, isolated, locations. The arrays can be used, for example, in high throughput genomics and proteomics for specific uses including, but not limited molecular diagnostics for early detection, diagnosis, treatment, prognosis, monitoring clinical response, and protein crystallography.

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.

The present invention includes a multivalent glycan microarray for detection of glycan-binding proteins. The multivalent glycan microarray allows a multivalent presentation of glycan on a microarray substrate, which can enhance binding of the glycan binding protein to the glycan microarray. The multivalent microarray includes a solid substrate having one or more branched polymers bonded to it via one or more silane-based linker reagents. The branched polymer in turn is bonded to a glycan, via one or more bifunctional linkers to form the multivalent glycan microarray. Nonspecific binding of glycan binding proteins to the multivalent glycan microarray can be reduced by using a blocking reagent coated on to the microarray substrate, which includes a polyethylene glycol surfactant attached to the solid substrate via a self-crosslinking azido-functionalized silane. Methods for making multivalent glycan microarrays and methods for using same to detect glycan-binding proteins are also disclosed.

The present invention includes a multivalent glycan microarray for detection of glycan-binding proteins. The multivalent glycan microarray allows a multivalent presentation of glycan on a microarray substrate, which can enhance binding of the glycan binding protein to the glycan microarray. The multivalent microarray includes a solid substrate having one or more branched polymers bonded to it via one or more silane-based linker reagents. The branched polymer in turn is bonded to a glycan, via one or more bifunctional linkers to form the multivalent glycan microarray. Nonspecific binding of glycan binding proteins to the multivalent glycan microarray can be reduced by using a blocking reagent coated on to the microarray substrate, which includes a polyethylene glycol surfactant attached to the solid substrate via a self-crosslinking azido-functionalized silane. Methods for making multivalent glycan microarrays and methods for using same to detect glycan-binding proteins are also disclosed.

De novo synthesized large libraries of nucleic acids are provided herein with low error rates. Further, devices for the manufacturing of high-quality building blocks, such as oligonucleotides, are described herein. Longer nucleic acids can be synthesized in parallel using microfluidic assemblies. Further, methods herein allow for the fast construction of large libraries of long, high-quality genes. Devices for the manufacturing of large libraries of long and high-quality nucleic acids are further described herein.