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Reusable plasmid extraction kit

A kit and plasmid technology, which is applied in the fields of molecular biology and pharmaceutical plasmid extraction, can solve the problems of complex preparation of plasmid extraction and purification columns, high cost of plasmid extraction kits, and non-reusable silica gel materials, etc., and achieves convenience in the plasmid extraction process Fast, high-purity extracted samples, and low-cost effects

Inactive Publication Date: 2013-04-17
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the high cost of the current plasmid extraction kit, the complicated preparation of the plasmid extraction and purification column, poor material mass transfer performance, limited utilization rate, low capacity, and the commonly used silica gel material cannot be reused, and the product is mainly imported and expensive, the present invention The purpose is to provide a kit for extracting plasmids with high purity and reusability, which can greatly reduce the operating cost of plasmid purification

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Synthesis of monolithic bed-type plasmid purification column: Weigh monomer glycidyl methacrylate, cross-linking agent trimerized isocyanurate triallyl urate (1:0.35mol / mol); initiator azobisisobutyronitrile (initiator agent / monomer 0.005:1mol / mol), porogen n-nonane (porogen / reaction mixture 70vol%), mix well, put into a plastic tube, and react at 50°C for 24 hours. Rinse the monolithic bed with ethanol. Then diethylamine is used as a modifier, ethanol is used as a solvent, the volume content of diethylamine is 75%, the modification reaction is controlled at 50°C, and the reaction time is 12 hours. After the reaction, wash with ethanol to obtain a monolithic bed type plasmid purification column.

Embodiment 2

[0068] Synthesis of monolithic bed type plasmid purification column: weigh monomer epoxy ethylene methacrylate, crosslinking agent glycerol trimethacrylate (1: 0.25mol / mol); initiator azobisisobutyronitrile (initiator porogen / monomer 0.03:1mol / mol); porogen ethylbenzene (porogen / reaction mixture 40vol%), mixed evenly, packed into a plastic tube, and reacted at 55°C for 20 hours. The monolithic bed was flushed with methanol. Then ethylamine was used as a modifier, methanol was used as a solvent, the volume content of ethylamine was 40%, the modification reaction was controlled at 70°C, and the reaction time was 16 hours. After the reaction, wash with ethanol to obtain a monolithic bed type plasmid purification column.

Embodiment 3

[0070] Synthesis of monolithic bed-type plasmid purification column: Weigh monomer glycidyl acrylate, cross-linking agent trimerized isocyanurate triallyl urate (1:0.28mol / mol); initiator azobisisoheptanonitrile (initiator / monomer 0.01: 1mol / mol); porogen xylene and n-octane (porogen / reaction mixture 50vol%, xylene / n-octane 1: 1vol / vol), mix well, put into a plastic tube, and React at 60°C for 16 hours. The monolithic bed was flushed with tetrahydrofuran. Then, ethylenediamine is used as a modifier, tetrahydrofuran is used as a solvent, the volume content of ethylenediamine is 60%, the modification reaction is controlled at 60° C., and the reaction time is 20 hours. After the reaction, wash with ethanol to obtain a monolithic bed type plasmid purification column.

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PUM

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Abstract

The invention discloses a reusable plasmid extraction kit, and relates to a plasmid extraction kit used in a nucleic acid extraction separation technique. The kit comprises a plasmid purification column, and series solution and thalli lysate for the plasmid purification column. The reusable plasmid extraction kit has the main technical characteristic that an integral bet-type plasmid purificationcolumn consisting of polymers and the series solution for the plasmid purification column are adopted. The integral bet-type plasmid purification column is simple to prepare, has high mass transfer property, strong pollution resistance, convenient and quick plasmid extraction process, and no obvious change of property after plasmid samples are extracted for many times. The reusable plasmid extraction kit overcomes the defects that the existing plasmid extraction kit has a high cost, and the plasmid purification column is complex to prepare, has poor material mass transfer property, limited utilization ratio, low volume and high price, and cannot be reused.

Description

technical field [0001] The invention relates to a plasmid extraction kit used in nucleic acid extraction and separation technology, in particular to a reusable kit capable of extracting and separating plasmids from a lysate and its application, belonging to the technical field of molecular biology and pharmaceutical plasmid extraction . Background technique [0002] Plasmid extraction is a common practice in the fields of molecular biology and pharmaceutical plasmid research. In recent years, gene therapy and DNA vaccines have shown outstanding advantages in the diagnosis and treatment of major diseases such as cancer, single-gene genetic diseases, AIDS, cardiovascular diseases, and arthritis, and their applied research is developing day by day. At present, plasmids have gradually become the most important gene delivery system for gene therapy, and the development of extraction technology for medicinal plasmids used in gene therapy has become one of the focuses of research ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 张敏莲靳琳陈然江骏
Owner TSINGHUA UNIV