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RNA extraction device and method

An extraction device and centrifuge tube technology, applied in the field of RNA extraction devices, can solve problems such as low binding efficiency of RNA sequences, degradation and destruction of RNA samples, reduction of proteinase K activity, etc., to improve purity, avoid sample loss and potential contamination, and reduce difficulties. degree of effect

Active Publication Date: 2020-05-29
SHANXI UNIV
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Problems solved by technology

Treat proteinase K at 100 degrees Celsius for 10 minutes, and DNaseI at 65 degrees Celsius for 10 minutes can be completely inactivated, but long-term high-temperature heating will also lead to degradation and destruction of RNA samples; phenol-chloroform extraction is the most stringent removal and inactivation The method of proteinase K and DNase I, but this method is cumbersome and time-consuming; although the EDTA method can inactivate DNase I and reduce the activity of proteinase K by chelating metal ions, EDTA may chelate metal ions and affect subsequent reverse transcription and PCR reactions
Although RNA purification methods such as silica gel binding and magnetic bead binding can use silica gel columns and magnetic beads to bind RNA within a certain length, and then remove most of proteinase K and DNase I through the eluent, the surface of the silica gel column and the magnetic A small amount of proteinase K and DNaseI will remain on the surface of the beads, and the binding efficiency of conventional silica gel columns and magnetic beads to ultra-long and ultra-short RNA sequences is low, which interferes with the results of subsequent precision experiments

Method used

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Embodiment Construction

[0039] In order to further illustrate the technical solution of the present invention, the present invention will be further described below through examples.

[0040] Such as Figure 1 to Figure 3 Shown, a kind of RNA extracting device comprises piston type syringe 1, mixing centrifuge tube 5, plastic centrifuge tube 2 and the spin column 3 that can be put into plastic centrifuge tube 2, is arranged on the needle outer surface of described piston type syringe 1 There is a ventilation groove 101, and a filter layer 4 is fixedly arranged in the spin column 3. The spin column 3 is further divided into a microporous ceramic filter layer spin column and a nitrocellulose membrane spin column according to different materials of the filter layer. A pressing plate can also be arranged on the filter layer 4 to compress the filter layer 4 so that the filter layer 4 is in close contact with the inner wall of the spin column 3 to prevent liquid leakage from the side wall gap. The filter ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to an RNA extraction device and method. The device comprises a piston type injector, a plastic centrifugal tube, a centrifugal column arranged in the plastic centrifugal tube and a mixing centrifugal tube, wherein the mixing centrifugal tube comprises a tube body and a tube cover; a through hole is formed in a pipe cover; a silica gel column is arranged in the through hole; a top separation layer is arranged at the upper part of an inner cavity of a pipe body; a middle separation layer is arranged at the middle-lower part of the inner cavity of the pipe body; the inner cavity of the pipe body is divided into an upper cavity, a middle cavity and a lower cavity by the top separation layer and the middle separation layer; grinding balls are pre-placed in the upper cavity; TRIzol is pre-placed in the middle cavity; and chloroform is pre-placed in the lower cavity. According to the invention, an operator doesnot need to directly suck and contact an organic solvent in the RNA extraction process, and toxic volatile matters are not released into an atmospheric environment in the operation process, so the health of experimenters is protected; and meanwhile, mechanical cutting and grinding of blocky tissue samples and difficult-to-split tissue samples are realized through the grinding balls, so RNA is released more quickly and fully.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an RNA extraction device and method. Background technique [0002] RNA, also known as ribonucleic acid Ribonucleic Acid, is one of the key substances in the life process. Extracting highly pure and intact RNA helps researchers to fully understand the regulatory mechanisms in the process of RNA transcription, splicing and degradation. At present, RNA extraction mainly includes silica gel binding method, magnetic bead binding method, CTAB or PVP method, cesium chloride density gradient centrifugation method, etc. Among them, the TRIzol method is one of the most widely used RNA extraction methods at present. This method has significant advantages such as large RNA yield, low cost, and good versatility. It is widely used in different tissues and tissues from animals, plants, fungi, bacteria, and viruses. Extract RNA from cells. [0003] TRIzol and chloroform are key reagen...

Claims

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Application Information

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IPC IPC(8): C12M1/42C12M1/00C12N15/10
CPCC12N15/1013C12N15/1017C12Q2521/537C12Q2521/119
Inventor 张学尧
Owner SHANXI UNIV
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