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Method for storing information in vivo by using DNA

A technology for internal storage and encoding of information, applied in recombinant DNA technology, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of high synthesis and storage costs, difficult storage, short storage time, etc., to achieve low cost, Improved storage capacity and low cost

Inactive Publication Date: 2020-01-14
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, the object of the present invention is to provide a method for storing information in the body using DNA in order to solve the problems in the prior art of using enzymes to catalyze the synthesis in vitro and store it with high cost, relatively difficult storage, and short storage time

Method used

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  • Method for storing information in vivo by using DNA
  • Method for storing information in vivo by using DNA
  • Method for storing information in vivo by using DNA

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, PCR preparation insert fragment and carrier

[0034] 1. Preparation of insert fragments

[0035] Q5 Master pool concentration: 14.024ng / μl

[0036] Table 1 insert

[0037] number of molecules copies per molecule Copy number corresponds to quality 12000 10 5

0.233ng

[0038] Dilute the master pool 200 times to a concentration of 0.07ng / μl.

[0039] The reaction system for PCR amplification using Q5 hot-start ultra-fidelity DNA polymerase is shown in Table 2.

[0040] Table 2 reaction system

[0041]

[0042]

[0043] The sequences of the forward and reverse primers are shown in Table 6.

[0044] The reaction program of PCR amplification is shown in Table 3.

[0045] Table 3 Reaction program

[0046]

[0047] After the PCR product is purified and recovered, use 30 μl ddH 2 O was dissolved, and the concentration of insert fragments was obtained by grayscale analysis.

[0048] 2. Preparation of carrier fragments ...

Embodiment 2

[0059] Embodiment 2, recombinant plasmid construction

[0060] 1, 3 or 5 inserts with corresponding homology arms were ligated with pUC19 vector using Gibson assembly kit, and reacted at 50°C for 60 min. The specific reaction system is shown in Table 7-9 below.

[0061] Table 7 The reaction system (pUC19-assembly-1) in which an insert fragment is connected to the pUC19 vector

[0062] components Dosage (μl) pUC19 linearized vector 2μl (200ng) insert fragment 1 1μl (13ng) Gibson Assembly Master Mix(2×) 10μl wxya 2 o

7μl

[0063] Table 8 The reaction system (pUC19-assembly-3) in which three inserts were connected to the pUC19 vector

[0064] components Dosage (μl) pUC19 linearized vector 2μl (200ng) insert fragment 1 1μl (13ng) insert fragment 2 1μl (13ng) insert fragment 3 1μl (13ng) Gibson Assembly Master Mix(2×) 10μl wxya 2 o

5μl

[0065] Table 9 The reaction syste...

Embodiment 3

[0067] Embodiment 3, recombinant plasmid transformation and cultivation

[0068] The plasmid assembled by Gibson was electrotransformed into E. coli competent cells, and the specific operations were as follows:

[0069] 1. Insert an electric cup with an electrode spacing of 0.1cm into crushed ice, and let it stand in the ice for 5 minutes to fully cool down the electric cup.

[0070] 2. Take the electroporation competent cells (Escherichia coli DH10β) stored at -70°C, the transformation efficiency is greater than 10 10 cfu / μgDNA) into the ice, after the cells have just thawed, add 5 μl of the ligated recombinant plasmid, immediately insert into the ice, and quickly transfer the cell / DNA mixture to the electric shock cup with a sterile tip in the ultra-clean bench to avoid Bubble, make sure the cells sink to the bottom of the cup, cover the cup, and keep the empty tube for later use.

[0071] 3. Start the electrorotator and set the shock parameters: 1.8kV. Put the electric c...

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Abstract

The invention belongs to the field of bioinformatics, and discloses a method for storing information in vivo by using DNA. The method includes: using PCR to amplify target fragments with different homology arms from a gene library with encoding information, assembling these fragments onto a plasmid vector using Gibson, transforming the plasmid vector into a microorganism for storage. The method uses DNA to encode the stored information in vivo, can successfully assemble different DNA fragments for encoding information by Gibson assembly on the carrier, and realize storage of digital information in vivo, and has advantages of low cost, easy storage and long storage time. In addition, because the information to be stored is cloned into the plasmid through Gibson assembly, the plasmid can bestably inherited in the presence of resistance, so that the information will not be lost, can be stably stored in a cell, and can be extracted when needed.

Description

technical field [0001] The invention belongs to the field of bioinformatics, and in particular relates to a method for storing information in vivo by using DNA. Background technique [0002] As the name implies, digital information is the digitalization of information. It is not a carrier of paper or other forms, but a high-tech information means that is composed of digital codes and uses Internet or other transmission channels as carriers. In 2016, humans generated a total of 16.1 trillion gigabytes of digital information; by 2025, this number is expected to increase more than tenfold. At present, the places where data are placed are mainly external hard disks and cloud server rooms, but most of them are not perfect solutions. Because they take up a lot of space and need to be upgraded every ten years or so. [0003] Compared with traditional electronic media, DNA has the characteristics of large capacity, small size, encoding, stable properties, long life and easy storag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/64C12N1/21C12N1/19C12R1/19
CPCC12N15/64
Inventor 齐浩
Owner TIANJIN UNIV
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