Abstract: Disclosed are methods for synthesizing and/or assembling at least one polynucleotide product having a predefined sequence from a plurality of different oligonucleotides. In exemplary embodiments, the methods involve synthesis and/or amplification of different oligonucleotides immobilized on a solid support, release of synthesized/amplified oligonucleotides in solution to form droplets, recognition and removal of error-containing oligonucleotides, moving or combining two droplets to allow hybridization and/or ligation between two different oligonucleotides, and further chain extension reaction following hybridization and/or ligation to hierarchically generate desired length of polynucleotide products.

Abstract: Genetically engineered microorganisms are provided that produce products from the fatty acid biosynthetic pathway (fatty acid derivatives), as well as methods of their use.

Abstract: The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer.

Abstract: The invention provides methods and compositions relating to cells having altered functions, and the nucleic acids that impart those functions. Altered cellular function arises from in vivo directed recombination of genetic elements to yield a recombined nucleic acid. These methods and compositions may utilize altered host cells having altered recombination enzyme profiles and/or altered recombination sites. The invention involves in some aspects methods for assembling nucleic acid molecules, such as genomic DNA. Aspects of the invention also provide kits, compositions, devices, and systems for generating novel recombined nucleic acids and cells having altered cell function.

Abstract: Certain aspects of the present invention provide artificial polypeptides containing one or more unnatural amino acids and methods for producing and using the artificial polypeptides.

Abstract: The invention relates to a method for detecting a double-stranded region in a nucleic acid by (1) providing two separate, adjacent pools of a medium and a interface between the two pools, the interface having a channel so dimensioned as to allow sequential monomer-by-monomer passage of a single-stranded nucleic acid, but not of a double-stranded nucleic acid, from one pool to the other pool; (2) placing a nucleic acid polymer in one of the two pools; and (3) taking measurements as each of the nucleotide monomers of the single-stranded nucleic acid polymer passes through the channel so as to differentiate between nucleotide monomers that are hybridized to another nucleotide monomer before entering the channel and nucleotide monomers that are not hybridized to another nucleotide monomer before entering the channel.

Abstract: The present invention provides Modular Molecular Clasps and methods of using these Modular Molecular Clasps in a wide range of applications in the health care industry, e.g., in therapy, in clinical diagnostics, in in vivo imaging or in drug discovery. The Modular Molecular Clasps of the present invention also have industrial and environmental applications, e.g., in environmental diagnostics, industrial diagnostics, food safety, toxicology, catalysis of reactions or high-throughput screening; as well as applications in the agricultural industry and in basic research. Methods of designing the Modular Molecular Clasps, as well as arrays and biosensors comprising these Modular Molecular Clasps are also provided.

Abstract: Disclosed are methods of manufacturing synthetic DNAs, that is, DNAs made at least in significant part by chemical synthesis of nucleic acid polymers. Also provided are methods for assembling plural DNAs in the same pool by multiplexed assembly of synthetic oligonucleotides. In exemplary embodiments, the methods involve pre-amplification of one or more oligonucleotides using “universal” primers, reduction of the error rate in oligonucleotide and/or nucleic acid products, and sequence optimization and oligonucleotides design. Also provided are low-purity arrays of nucleic acids and methods for assembling nucleic acids using oligonucleotides obtained from low-purity arrays.

Abstract: The present invention relates to methods, cells and compositions for performing in vivo recombination without the need for clonal selection. These methods, cells and compositions are useful for the automation of recombination. The present invention provides methods for performing sequential steps in genome engineering, as well as for achieving parallel and multiplex genome engineering.

Abstract: Novel methods and compositions for DNA sequencing are provided. The methods described herein are useful for sequencing homopolymeric regions of DNA. The methods also prevent the accumulation of mistakes and inefficiencies in the sequencing reaction.

Abstract: Compositions and methods for controlling and tracking items are provided. More particularly, labeling compositions useful as a coding system to deter diversion of items of medication or other valued items are provided, wherein the labeling compositions comprise at least 25 unique identifiers, wherein the at least 25 unique identifiers are selected from a set of at least 50 unique identifiers. Also provided are methods for using such compositions to provide trackability throughout a chain of custody.

Abstract: Disclosed are methods of manufacturing synthetic DNAs, that is, DNAs made at least in significant part by chemical synthesis of polynucleotide polymers. Also provided are methods for assembling plural DNAs in the same pool by multiplexed assembly of synthetic oligonucleotides. In exemplary embodiments, the methods involve pre-amplification of one or more oligonucleotides using “universal” primers, reduction of the error rate in oligonucleotide and/or polynucleotide products, and sequence optimization and oligonucleotides design.

Abstract: Miniaturized, high-density, bead-based arrays are provided. Methods of producing and using clonal beads and producing and using miniaturized, high density, bead-based arrays are also provided.

Abstract: The invention relates to a method for detecting a double-stranded region in a nucleic acid by (1) providing two separate, adjacent pools of a medium and a interface between the two pools, the interface having a channel so dimensioned as to allow sequential monomer-by-monomer passage of a single-stranded nucleic acid, but not of a double-stranded nucleic acid, from one pool to the other pool; (2) placing a nucleic acid polymer in one of the two pools; and (3) taking measurements as each of the nucleotide monomers of the single-stranded nucleic acid polymer passes through the channel so as to differentiate between nucleotide monomers that are hybridized to another nucleotide monomer before entering the channel and nucleotide monomers that are not hybridized to another nucleotide monomer before entering the channel.

Abstract: The present invention provides recombination based methods for assembling nucleic acids. In certain aspects the present invention provides hierarchical assembly methods for producing genome sized polynucleotide constructs. The methods may be used for assembling large polynucleotide constructs, for synthesizing synthetic genomes, or for introducing a plurality of nucleotide changes throughout the genome of an organism. In another aspect, the invention provides cells having increased genomic stability. For example, cells comprising alterations in at least a substantial portion of the transposons in the genome are provided.

Abstract: Disclosed are methods of assembling nucleic acid constructs from component parts in a manner that is not dependent on the sequence of the component parts. The methods may be used to assemble large nucleic acid constructs from multiple component parts in one or more reactions. The methods may also be used to assemble two or more nucleic acid constructs in the same reaction mixture. In exemplary embodiments, the methods involve formation of a Holliday junction or bridge structure using a junction oligonucleotide.

Abstract: Disclosed are methods of manufacturing synthetic DNAs, that is, DNAs made at least in significant part by chemical synthesis of polynucleotide polymers. Also provided are methods for assembling plural DNAs in the same pool by multiplexed assembly of synthetic oligonucleotides. In exemplary embodiments, the methods involve pre-amplification of one or more oligonucleotides using “universal” primers, reduction of the error rate in oligonucleotide and/or polynucleotide products, and sequence optimization and oligonucleotides design.

Abstract: In certain aspects the present invention provides methods and compositions related to rational protein design.

Abstract: Methods of improving the kinetics of bimolecular interactions where reactants are present in low concentrations are provided. Methods of pre-amplifying one or more oligonucleotides using high concentration universal primers are provided. Methods of improving the error rate in oligonucleotide and/or polynucleotide syntheses are also provided. Methods for sequence optimization and oligonucleotides design are further provided.

Abstract: Methods of reducing background during amplification of small amounts of nucleic acids employ careful analysis of sources of low level contamination. Ultraviolet light can be used to reduce nucleic acid contaminants in reagents and equipment. “Primer-dimer” background can be reduced by judicious design of primers. We have shown clean signal-to-noise with as little as starting material as one single human cell (˜6 picogram), E. coli cell (˜5 femtogram) or Prochlorococcus cell (˜3 femtogram).