CN106170547 Method for preparing DNA unit composition, and method for creating concatenated DNA

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Application Number 201480073826.9

Application Date 05.09.2014

Publication Number 106170547

Publication Date 30.11.2016

Grant Number 106170547

Grant Date 27.03.2020

Publication Kind B

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Applicants TECHNOLOGY RESEARCH ASSOCIATION OF HIGHLY EFFICIENT GENE DESIGN
株式会社辛普罗根

Inventors TSUGE KENJI
柘植谦尔
ITAYA MITSUHIRO
板谷光泰

Agents 北京市金杜律师事务所 11256
北京市金杜律师事务所 11256

Priority Data 2014-008690 21.01.2014 JP

Title

(EN) Method for preparing DNA unit composition, and method for creating concatenated DNA

(ZH) 单元DNA组合物的制备方法及DNA连结体的制作方法

Abstract

(EN) Provided are: a method for preparing a DNA unit composition in which the mol number of a plurality of DNA units is more uniform, and a method for creating concatenated DNA. The method for preparing a DNA unit composition has: a step for preparing solutions which contain a plurality of DNA units to which an added sequence is linked, and preparing a solution for each type of DNA unit; and a step for, after preparing each of the solutions, measuring the concentration of the DNA unit in each of the solutions in a state where the added sequence is linked to the DNA unit, and on the basis of the results thereof, fractionating each of the solutions and making the mol number of the DNA unit in each of the solutions closer to being identical to one another. The method for creating concatenated DNA has: a step for preparing the DNA unit composition; a step for preparing a vector DNA; a step for removing each of the added sequences, by using a restriction enzyme, from the DNA units to which the added sequence is linked in the solutions after preparation thereof; and a step for, after the removal step, linking the vector DNA and each of the DNA units with one another.

(ZH) 本发明提供多个单元DNA的摩尔数更加一致的单元DNA组合物的制备方法及DNA连结体的制作方法。单元DNA组合物的制备方法包括下述工序：准备各自含有连结有附加序列的多个单元DNA中的每一种所述单元DNA的溶液的工序；和，在准备各溶液后，在单元DNA连结有附加序列的状态下，测定各溶液中的单元DNA的浓度，基于其结果分取各溶液，使各溶液中的单元DNA的摩尔数接近于彼此相同的工序。DNA连结体的制作方法包括下述工序：制备单元DNA组合物的工序、准备载体DNA的工序、用限制性内切酶从制备得到的溶液中的连结有附加序列的单元DNA中除去各附加序列的工序、和在除去工序后将载体DNA及各单元DNA彼此连结的工序。

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12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology

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12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
10	Processes for the isolation, preparation or purification of DNA or RNA

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
63	Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
64	General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host

C	CHEMISTRY; METALLURGY
12	BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N	MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15	Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09	Recombinant DNA-technology
63	Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
66	General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease

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1. CN106170547 - Method for preparing DNA unit composition, and method for creating concatenated DNA