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Device for gene purification and purification method

A gene and formula technology, applied in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problems of high cost, cumbersome operation, long extraction time, etc., and achieve simple operation and time saving. , cost saving effect

Inactive Publication Date: 2016-07-13
ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the existing problems in the process of gene purification, such as the use of toxic organic solvents, repeated use of centrifuges, cumbersome operation, long extraction time, and high cost, the present invention provides a gene purification method

Method used

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  • Device for gene purification and purification method
  • Device for gene purification and purification method
  • Device for gene purification and purification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] A kind of device structure for gene purification of the present invention is as follows:

[0088] Such as figure 1 As shown, the device for bacterial gene purification of the present invention is based on a common syringe, adding a sieve plate and an adsorption material. The specific structure is as follows: the device of the present invention includes a suction head 4, a sleeve 1, a sieve plate 3 and a push rod 2, the suction head 4 is closely attached to the front end of the sleeve 1, and the sieve plate 3 is matched with the cross-sectional size of the inner wall of the sleeve Disc shape, the sieve plate 3 and the push rod 2 are sequentially placed inside the sleeve 1, and are closely attached to the inner wall of the sleeve 1 respectively. Between the sieve plate 3 and the front end of the sleeve 1, an adsorption material, such as SiO with a particle size of 20-80 μm, is placed 2 powder.

[0089] The volume of the device vs. SiO 2 The dosage ratio is: 2.5ml: 0.1...

Embodiment 2

[0093] A reagent for extracting bacterial plasmid DNA, consisting of the following components:

[0094] Cell suspension, cell lysate, plasmid binding solution, deproteinization solution, washing solution, eluent.

[0095] Wherein, the formula of the cell suspension is: 50mmol / LTris, 10mmol / LEDTA, 1-10μg / mL RNaseA; ultrapure water is used as solvent; pH6.0-9.0, stored at 4°C.

[0096] The formula of the cell lysate is: 0.05-1.5mol / L NaOH, SDS with a mass volume percentage of 1%-3%; ultrapure water is used as a solvent, and it is stored airtight at room temperature. When the above-mentioned components and component concentrations are used, the lysis is mild and rapid, with little damage to the genome and plasmid, which helps to reduce genome pollution and protect the supercoiled structure of the plasmid.

[0097] The formula of the plasmid binding solution is: 2-6mol / L GuHCl, 0.01-1mol / LCH 3 COOK; use ultrapure water as solvent, pH 3-5, store in airtight at room temperature. ...

Embodiment 3

[0115] A reagent for extracting bacterial genomic DNA, consisting of the following components:

[0116] Cell suspension, cell lysate, neutralization solution, deproteinization solution, washing solution, eluent.

[0117] Among them, the formula of the cell suspension is: 50mmol / LTris, 10mmol / LEDTA, 1μg / mL RNaseA; use ultrapure water as solvent; pH6.0-9.0, store at 4°C.

[0118] The formulation of the cell lysate is: TritonX-100 with a volume percentage of 2%-6%, using ultrapure water as a solvent, and storing in airtight at room temperature. TritonX-100 with a volume percentage of 2%-6% is mildly lysed and has no damage to the genome.

[0119] The formula of the neutralizing solution is: 2-6mol / L GuHCl, 0.5-2mol / LLiCl, 30%-60% ethanol by volume; ultrapure water is used as solvent, pH 5-7, and airtight storage at room temperature. After adding the neutralizing solution, the solution has no flocculent precipitation, no need for shaking, and can better maintain the integrity of...

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Abstract

The invention provides a device for gene purification. The device comprises a sucker, a sleeve, a screening plate and a push rod, wherein the sucker is tightly adhered to the front end of the sleeve; the screening plate takes the shape of a circular piece which is matched with the cross section of the inner wall of the sleeve in size; the screening plate and the push rod are sequentially arranged inside the sleeve and are tightly adhered to the inner side wall of the sleeve; an absorbing material is placed between the screening plate and the front end of the sleeve. The invention further provides a method for gene purification by using the device. Low-cost and high-efficiency purification of bacterial genome and plasmid, PCR Clean and rubber recycling can be achieved by using normal consumables and medicines of laboratories, and moreover no toxic organic solvent is involved in the whole extraction process. The device provided by the invention is simple in manufacturing method and operation method and can be recycled, and the cost can be lowered. No centrifuge is used in the washing and elution process, and ethanol is not needed to be volatized before elution, so that the time can be shortened. The device is rich in specification, and trace and large-scale operation can be implemented.

Description

technical field [0001] The invention relates to a device and a purification method for gene purification. Background technique [0002] Plasmid is used as an important carrier for carrying foreign genes into bacteria for amplification or expression. It can introduce target DNA fragments into recipient cells for replication and expression through recombinant DNA technology. Therefore, the quality of plasmid extraction plays a key role in the success or failure of subsequent molecular biology experiments. At present, many domestic and foreign companies have developed and provided plasmid extraction kits, but when these kits extract plasmids, they need to go through cumbersome steps and take a long time. And many poisonous chemical substances are adopted in the test kit, which will pollute the environment and threaten the health of the operator. [0003] At present, the methods for extracting bacterial plasmid DNA mainly include: 1. Phenol-chloroform-isoamyl alcohol extractio...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12N15/10
CPCC12N15/101C12Q2523/308C12Q2527/125C12Q2521/537
Inventor 薛正莲杨建伟朱昊王洲刘艳费芙蓉吴爽彭凡袁红梅
Owner ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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