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Fidelity storage method for yeast DNA (Deoxyribonucleic Acid)

A yeast and fidelity technology, applied in the field of DNA material storage research, can solve the problems of low fidelity, unrealistic large-scale information storage, inconvenient access, etc., achieve high fidelity, solve large-scale information storage Effects with little problem, impact or damage

Inactive Publication Date: 2019-03-01
EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This preservation method will affect or damage the DNA, the fidelity is not high, and it is not realistic in large-scale information preservation, and it is not convenient for frequent access

Method used

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  • Fidelity storage method for yeast DNA (Deoxyribonucleic Acid)
  • Fidelity storage method for yeast DNA (Deoxyribonucleic Acid)
  • Fidelity storage method for yeast DNA (Deoxyribonucleic Acid)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0014] 1. Bead milling method to extract the DNA in the sample: take 4ml of active yeast liquid in a 5ml centrifuge tube, centrifuge at 8000g for 6min, resuspend the bacteria with 1600ul STE buffer, and wash the bacteria by centrifugation twice under the same conditions; the collected bacteria After suspending with 800ul TE buffer, add about 100mg of glass beads with a diameter of 425-600um, and then add 400ul Tirs saturated phenol; the above mixture is shaken on a vortex shaker for 300s, then placed in a refrigerator at 4°C for 6min, centrifuged at 13000g 6min; transfer the upper aqueous phase (about 640ul) to a new centrifuge tube, add TE buffer to make up to 800ul, then add 400ul chloroform, mix well, refrigerate at 4°C for 5min, centrifuge at 13000g for 10min. Repeat this step two to three times; transfer the upper aqueous phase (about 640ul) to a new centrifuge tube, add 200ul TE and 10ul RNase (10mg / ml), digest at 37°C for 13min; add 400ul chloroform to mix, and incubate ...

experiment example

[0017] 1. Method:

[0018] 1. Quantitative measurement of extracted DNA by ultraviolet absorption method: take 2 centrifuge tubes, add the DNA nucleic acid solution extracted in step a) respectively, then add 2mL distilled water (as a blank control) to tube A, and add 2mL molybdic acid to tube B Ammonium-perchloric acid precipitant mixed evenly;

[0019] 2. Place in an ice bath (or refrigerator) for 30 minutes, centrifuge at 3000r / min for 10 minutes; dilute the supernatants of tubes A and B respectively until the optical density is between 0.1 and 1.0;

[0020] 3. Select a quartz cuvette with an optical path of 1 cm, and at a wavelength of 260 nm, its optical density OD260nm can be calculated according to the Lambert-Beer theorem (Formula 1), and the DNA content can be calculated according to Formula 2.

[0021]

[0022]

[0023] Among them, l is the thickness of the light passing through the object to be measured, and d is the dilution factor.

[0024] 2. Exploration ...

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Abstract

The invention discloses a fidelity storage method for yeast DNA (Deoxyribonucleic Acid). The fidelity storage method comprises the following steps: enabling a yeast sample solution to be thallus re-suspension with TE buffer solution; centrifuging and cleaning; adding glass beads and Tirs saturated phenol to obtain a mixed solution, oscillating the mixed solution for 5 to 10 minutes, refrigeratingand centrifuging; absorbing an upper-layer water phase and transferring the upper-layer water phase to a new centrifuging tube; adding TE and RNase for digestion, adding chloroform, uniformly mixing,refrigerating and centrifuging; transferring the upper-layer water phase to a new centrifuging tube; storing under ultraviolet rays at a temperature of 15 to 25 DEG C, and at pH of 7 to 8. By adoptingthe fidelity storage method, in the extraction of the DNA, the mixed solution is oscillated for 5 to 10 minutes, and is stored under the irradiation of the ultraviolet rays at the temperature of 15 to 25 DEG C, and the pH of 7 to 8, so that the influence or damage on the DNA is lowest, highest fidelity rate is achieved, and the problem concerned with large-scale information storage is solved.

Description

technical field [0001] The invention relates to the technical field of DNA material storage research, in particular to a fidelity storage method of yeast DNA. Background technique [0002] The original biological computing technology is to use the arrangement and combination of massive DNA sequences to solve NP-hard problems. Its core technology is based on the characteristics of a large number of DNA nucleotides in a small volume, using the characteristics of pairing between DNA nucleotides, and solving large-scale permutation and combination problems through the method of exhaustive pairing of nucleotides, so as to solve the problem of NP -hard question purpose. The new generation of biological computing technology has involved the construction and integration of basic computing and logic modules. The two most notable directions are electronic cells and synthetic biology: the concept of electronic cells is based on the in-depth understanding of the expression and regulat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/10C12N15/1006
Inventor 宁康李锐豪陈超云朱雪
Owner EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH
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