Preparation method of long single-chain DNA

A single-strand, sequence technology, applied in the field of preparation of long single-strand DNA, can solve the problem that the single-strand DNA sequence cannot be customized, and achieve the effect of efficient cutting, high-efficiency preparation, and high purity
CN110699407APending Publication Date: 2020-01-17FUNDAN UNIVERSITY SHANGHAI CANER CENTER
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
FUNDAN UNIVERSITY SHANGHAI CANER CENTER
Publication Date
2020-01-17

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Abstract

The present invention discloses a method for efficiently preparing long single-chain DNA by means of class I and class II hydrolysable deoxyribozymes. The method mainly comprises steps of designing and constructing recombinant phagemids to obtain phagemid circular single-chain DNA, at the same time, the class I deoxyribozyme and class II deoxyribozyme mutants are used to cut the circular single-chain DNA, and the long single-chain DNA is obtained by purifying, recovering and enzyme digestion. The two types of the deoxyribozymes capable of rapidly hydrolyzing DNA are used to replace restrictionendonucleases, the preparation method realizes specific cleavage of the prepared DNA sequences by an assisted phage method in a low cost manner, and the single-chain DNA with any length and sequencesis prepared in a large amount, economic and high-purity manner.
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Description

technical field

[0001] The invention belongs to the fields of biochemistry and molecular biology, and specifically relates to a method for preparing long single-stranded DNA by using two types of deoxyribozymes capable of rapidly hydrolyzing DNA and combining with helper phages. Background technique

[0002] At present, DNA nanotechnology, gene editing such as knock in (gene knock-in) and other biomedical research fields have a wide demand for single-stranded DNA (Single strand DNA, ssDNA), especially for long single-stranded DNA (>100 bases). However, limited by chemical synthesis methods, the synthesis of long single-stranded DNA is difficult to guarantee yield, yield and satisfactory cost performance. Therefore, for long single-stranded DNA, it is necessary to rely on the in vivo or in vitro action of biological enzymes and some means of assisted denaturation. Commonly used methods for preparing long single-stranded DNA mainly include reverse transcription, enzymatic d...

Claims

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