The present invention provides methods, compositions, and systems for distributing single polymerase molecules into array regions. In particular, the methods, compositions, and systems of the present invention result in a distribution of single polymerase molecules into array regions at a percentage that is larger than the percentage expected to be occupied under a Poisson distribution.

This disclosure demonstrates an approach that translates synthetic DNA codes to spatial codes registered in nanoliter microchambers for multiplexed measurement of nearly any type of molecular targets (e.g., miRNAs, mRNAs, intracellular and surface proteins) in single cells.

This disclosure demonstrates an approach that translates synthetic DNA codes to spatial codes registered in nanoliter microchambers for multiplexed measurement of nearly any type of molecular targets (e.g., miRNAs, mRNAs, intracellular and surface proteins) in single cells.

This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.

This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.

Nucleobase amino acid polymer compositions and methods for synthesizing nucleobase amino acid polymers and their conversion to nucleic acids are provided. The nucleobase amino acid polymer compositions disclosed herein comprise nucleobase amino acid monomers that comprise a linker and a nucleobase. The methods disclosed herein comprise synthesizing a nucleic acid polymer by providing a mRNA template, at least one ribosome, at least one nucleobase amino acid tRNA and at least one non-nucleobase amino acid tRNA; and adding a polymerase and at least one primer to the nucleobase amino acid polymer.

Nucleobase amino acid polymer compositions and methods for synthesizing nucleobase amino acid polymers and their conversion to nucleic acids are provided. The nucleobase amino acid polymer compositions disclosed herein comprise nucleobase amino acid monomers that comprise a linker and a nucleobase. The methods disclosed herein comprise synthesizing a nucleic acid polymer by providing a mRNA template, at least one ribosome, at least one nucleobase amino acid tRNA and at least one non-nucleobase amino acid tRNA; and adding a polymerase and at least one primer to the nucleobase amino acid polymer.

The present disclosure relates to tagged nucleoside-5-oligophosphates having a positively charged polymer tag structure and components thereof. Such nucleoside-5-oligophosphates are useful, for example, in nanopore-based sequencing-by-synthesis applications. Also disclosed herein are nanopore constructs engineered to have additional negatively-charged moieties in the channel of the nanopore. Such nanopores can be useful, for example, for providing a repellant force against template and/or primer nucleic acids inserting into the pore during a nucleic sequence-by-synthesis process. The tagged nucleoside-5-oligophosphates and nanopores disclosed herein can be used together to provide nanopore-based nucleic acid sequencing-by-synthesis systems and processes having reduced background tag levels and improved throughput.

The present disclosure relates to tagged nucleoside-5-oligophosphates having a positively charged polymer tag structure and components thereof. Such nucleoside-5-oligophosphates are useful, for example, in nanopore-based sequencing-by-synthesis applications. Also disclosed herein are nanopore constructs engineered to have additional negatively-charged moieties in the channel of the nanopore. Such nanopores can be useful, for example, for providing a repellant force against template and/or primer nucleic acids inserting into the pore during a nucleic sequence-by-synthesis process. The tagged nucleoside-5-oligophosphates and nanopores disclosed herein can be used together to provide nanopore-based nucleic acid sequencing-by-synthesis systems and processes having reduced background tag levels and improved throughput.

The invention provides compositions, including tethered synaptic complexes (TSCs), artificial nucleic acids, molecular constructs that include artificial nucleic acids bound to transposases, and kits; as well as methods of using the same, for example, for preparation of nucleic acid libraries and sequencing.