34 results about "Oligonucleotide Linker" patented technology

The invention discloses an oligonucleotide-linker-mediated DNA assembly method and an application thereof. The assembly method comprises steps of synthesis of a long sequence, preparation and assembly of a module carrier, remolding and assembly of a skeleton carrier, preparation of a short sequence, and enzyme-cut and link-up integration of the module carrier, the remolded skeleton carrier and the short sequence. According to the method, the short sequence is a regulating element such as a promoter and an RBS, and the long sequence is a replication initiation point of a gene or a carrier. The short sequence is designed to synthesize a double-stranded DNA chain, and can connect the long sequence and be used as a barcode sequence to determine the assembly sequence in an assembly process. The gene sequence in an operon is achieved by synthesizing short sequence with different cohesive ends. According to the method, multiple factors are allowed to participate in gene regulation at the same time. The method can be used to construct a combinatorial library of a biosynthesis channel, and a high-flux optimized channel is achieved.

The present invention provides a method for the assembly of a polynucleic acid sequence from a plurality of nucleic acid sequences in which the polynucleic acid sequence is of a formula Nn+1, in which N represents a nucleic acid sequence and where n is 1 or greater than 1 and each N may be the same or a different nucleic acid sequence, in which the method comprises: (i) providing a first nucleic acid sequence N1 which has an oligonucleotide linker sequence L13 at the 3′-end of the nucleic acid sequence; (ii) providing a second nucleic acid sequence N2 which optionally has an oligonucleotide linker sequence L23′ at the 3′-end of the nucleic acid sequence and which has an oligonucleotide linker sequence L25′ at the 5′-end of the nucleic acid sequence, wherein the 5′-end linker sequence L25′ of nucleic acid sequence N2 is complementary to the 3′-end linker sequence L13′ of nucleic acid sequence N1; (iii) optionally providing one or more additional nucleic acid sequences N, wherein nucleic acid sequence N2 has an oligonucleotide linker sequence L23′ at the 3′-end of the nucleic acid sequence, and wherein said one or more additional nucleic acid sequences N comprises a terminal additional nucleic acid sequence NZ, and wherein each additional nucleic acid sequence N has an oligonucleotide linker sequence at its 3′-end, wherein said terminal additional nucleic acid sequence NZ optionally lacks an oligonucleotide linker sequence at its 3′-end and wherein each additional nucleic acid sequence N has an oligonucleotide linker sequence at its 5′-end, wherein for the first additional nucleic acid sequence N3 the 5′-end linker sequence L35′ is complementary to the 3′-end linker sequence L23′ of nucleic acid sequence N2 and for each second and subsequent additional nucleic acid sequence N the 5′-end linker sequence is complementary to the 3′-end linker sequence of the respective preceding additional nucleic acid sequence; and (iv) ligating said nucleic acid sequences to form said polynucleic acid sequence.

Currently, the circularization of small RNAs is broadly regarded as an obstacle in ligation-related assays and explicitly avoided while short lengths of linear RNA targets is broadly recognized as a factor limiting use of conventional primers in PCR-related assays. In contrast, the disclosed invention capitalizes on circularization of small RNA targets or their conjugates with oligonucleotide adapters. The circular RNA templates provide amplification of the target sequences via synthesis of multimer nucleic acids that can be either labeled for direct detection or subjected to PCR amplification and detection. Structure of small circular RNAs and corresponding multimeric nucleic acids provide certain advantages over current methods including flexibility in design of conventional RT and PCR primers as well as use of 5′-overlapping dimer-primers for efficient and sequence-specific amplification of short target sequences. Our invention also reduces number of steps and reagents while increasing sensitivity and accuracy of detection of small RNAs with both 2′OH and 2′-OMe at their 3′ ends. Our invention increase sensitivity and specificity of detection of microRNAs and other small RNAs with both 2′OH and 2′-OMe at their 3′ ends while allowing us to distinguish these two forms from each other.

Oligonucleotide structures are provided that are capable of forming more stable bonds to a lipid membrane and thereby generate an improved control of the process whereby oligonucleotide linkers are introduced to lipid membranes. Methods of forming lipid membrane oligonucleotide attachments are provided including lipid vesicles. The oligonucleotides typically comprise at least two hydrophobic anchoring moieties capable of being attached to a lipid membrane. Said moieties may be attached at the terminal ends of an oligonucleotide or, in the case of a first and second strand forming a duplex, at the same terminal end one of the strands other end not being part of the duplex leaving it free to hybridize to additional strands. The lipid vesicles attached with the oligonucleotide can be used in biosensors and may contain membrane proteins.

The present invention provides a simple and rapid method for preparing purified transposase complexes that are highly suited for fragmenting DNA. The method includes forming transposase complexes with oligonucleotide adapters in cell lysate, then purifying the complexes from the other substance in the cell lysate. Purification is accomplished using a specific binding pair, in which one member of the pair is bound to an oligonucleotide adapter of the complex and the other member of the pair is bound to a solid substrate. The bound complexes can be immediately used in DNA fragmentation reactions to produce solid substrate-bound DNA fragments, which can be used for any number of purposes, including as templates for amplification and sequencing.

Controlled delivery of nucleic acid into target cells is achieved by immobilizing the nucleic acid and the cells to a substrate. Improved control over delivery is achieved by immobilizing the nucleic acid to the substrate via complementary DNA binding interactions with an oligonucleotide linker.

The invention discloses an antibody antigen-binding fragment-aplysiatoxin conjugate and a preparation method and application thereof. The preparation method includes steps: (1) providing an antigen-binding fragment of an anti-CD20 monoclonal antibody with an LPXTG sequence at a C-terminal and aplysiatoxin or derivatives thereof with oligonucleotide linkers; (2) under Sortase A enzyme catalysis, the LPXTG sequence and the oligonucleotide linkers have peptide transferase reaction to enable coupling of the antigen-binding fragment with the aplysiatoxin or derivatives thereof with the oligonucleotide linkers; (3) after reaction is completed, separating and purifying to obtain the antigen-binding fragment-aplysiatoxin conjugate. The antigen-binding fragment-aplysiatoxin conjugate has advantagesthat specificity, affinity and CD20 positive tumor cell killing effects of whole antigen-aplysiatoxin conjugates are retained, and high tumor penetrating rate and high tumor position clustering quantity are realized.