8 results about "Nuclease digestion" patented technology

The present invention relates to methods for producing a protected deoxyribonucleic acid (DNA) product with enhanced resistance to nuclease digestion. The methods comprise digesting a double-stranded DNA molecule with an endonuclease to generate a digested double-stranded DNA molecule comprising a first strand and a second strand, wherein the first strand has a 5' overhang; and extending the second strand 5' to 3' using a polymerase in the presence of one or more nuclease-resistant deoxynucleotide triphosphates to generate the protected DNA product, wherein the protected DNA product comprises an extended second strand comprising at least one nuclease-resistant nucleotide incorporated into the extended second strand by the polymerase using the 5' overhang of the first strand as template. Also provided are protected DNA products and uses thereof.

The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides employing hybridization stringency and / or nuclease digestion for removing failure sequences.

Linear DNA with improved resistance against exonucleases and methods for its production. The present invention relates to methods for producing a linear deoxyribonucleic acid (DNA) product (e.g., a closed linear DNA product) with improved resistance to nuclease digestion. The present invention relates to methods comprising the steps of: (a) contacting a double-stranded DNA molecule with an endonuclease, a ligase, and first and second terminal adaptor molecules and intermediate adaptor molecules to form a single contiguous aqueous volume; and (b) incubating the single contiguous aqueous volume to generate a linear DNA product (e.g., a closed linear DNA product). The present invention also relates to linear deoxyribonucleic acid (DNA) products (e.g., a closed linear DNA product) with improved resistance to nuclease digestion and uses thereof.

The present invention relates to methods for producing a linear deoxyribonucleic acid (DNA) product (e.g. a closed linear DNA product) with enhanced resistance to nuclease digestion using a method comprising a cas nuclease. The present invention relates to methods comprising contacting a doublestranded DNA molecule with an RNP complex comprising a single stranded guide RNA (gRNA) and a cas nuclease, a ligase and first and second adaptor molecules to generate a linear DNA product. The step of contacting the DNA molecule with the RNP complex may be carried out prior to or simultaneously with the step of contacting with a ligase and first and second adaptor molecules.

In certain embodiments, the present invention provides a detection composition comprising a picodroplet comprising (a) an aqueous solution, and (b) a substrate probe comprising (i) an oligonucleotide of 2 to 75 nucleotides in length, (ii) a fluorophore operably linked to the oligonucleotide, and (iii) a quencher operably linked to the oligonucleotide. As used herein, the term “picodroplet” comprises a liquid droplet that has a volume of 0.014 to 2.6 picoliters. In certain embodiments, the present invention provides a method of detecting at least one individual nuclease molecule present in a sample, comprising contacting an aqueous sample suspected of containing at least one nuclease with at least one detection composition comprising a picodroplet comprising (a) an aqueous solution, and (b) a substrate probe comprising (i) an oligonucleotide of 2 to 75 nucleotides in length, (ii) a fluorophore operably linked to the oligonucleotide, and (iii) a quencher operably linked to the oligonucleotide to form an aqueous reaction mixture; emulsifying the aqueous mixture in oil to form picoliter-scale droplets in an emulsion, (c) incubating the picoliter-scale droplets in the emulsion in order for the nuclease, if present, to digest the substrate probes linked to the microbeads; recovering the microbeads; and detecting fluorescence emitting from the microbeads.

The invention relates to methods for producing a linear deoxyribonucleic acid (DNA) product with enhanced resistance to nuclease digestion. The linear DNA product may be a closed linear DNA product, a linear DNA product comprising nuclease-resistant nucleotides, or a partially closed DNA product.

The present invention relates to methods for producing a linear deoxyribonucleic acid (DNA) product (e.g. a closed linear DNA product) with enhanced resistance to nuclease digestion using a method comprising a Cas nuclease. The present invention relates to methods comprising contacting a double-stranded DNA molecule with an RNP complex comprising a guide RNA (gRNA) (e.g. a single stranded gRNA) and a Cas nuclease, a ligase and first and second adaptor molecules to generate a linear DNA product. The step of contacting the DNA molecule with the RNP complex may be carried out prior to or simultaneously with the step of contacting with a ligase and first and second adaptor molecules.

The invention discloses an information storage, encryption and decryption method based on a DNA nano switch, and belongs to the technical field of biomolecule self-assembly material information storage and encryption. According to the method, a key sequence is protected by means of an RNA chain, formation of an annular DNA nano-switch is prevented, and information storage and encryption are achieved, a target chain Tx chain is added in advance, then a secret key RNaseA nuclease is added to digest the RNA protection chain, the target chain Tx and a detection chain Dx form the annular DNA nano-switch based on base complementary pairing, RNaseA can digest all the RNA protection chains in a solution, and the RNA protection chains in the solution can be separated from the target chain Tx. The method has the advantages that the RNA protection chain Rx and the detection chain Dx are connected to form the annular DNA nano switch Sx, secondary connection of the residual RNA protection chain Rx and the detection chain Dx is avoided, the target chain Tx and the detection chain Dx are connected to form the annular DNA nano switch Sx after RNAseA nuclease digests the RNA protection chain, the agarose gel imaging analysis technology is combined, rapid decryption and accurate reading of molecular-level information can be achieved, and the accuracy of information encryption is improved. And a basic strategy is provided for promoting the development of a nano-scale biological information encryption technology.