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Method for extracting RNA from animal cartilage tissue

A cartilage tissue and animal technology, applied in the field of RNA extraction, can solve the problems of high polysaccharide content and difficulty in RNA extraction, and achieve the effects of high extraction quantity and quality, low cost and short time-consuming.

Inactive Publication Date: 2018-02-16
SHANXI AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem of difficult RNA extraction due to the high content of polysaccharides in tibia, a method for extracting RNA from animal cartilage tissue is provided.

Method used

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  • Method for extracting RNA from animal cartilage tissue
  • Method for extracting RNA from animal cartilage tissue
  • Method for extracting RNA from animal cartilage tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Chicken tibia growth plate total RNA extraction

[0027] (1) Put 0.05 g of chicken tibial growth plate stored at -80°C into a mortar filled with liquid nitrogen, grind it thoroughly until it becomes powdery, then add 0.5 ml of Trizol reagent (Invitrogen, Tankara, etc.), and wait until Continue to grind thoroughly when it is converted into a solution, and the time is 1 min; transfer the solution in the mortar to a 1 ml centrifuge tube (RNase-free, sterile), mix vigorously, let stand for 5 min, add 100 ul chloroform, shake vigorously to mix Evenly, let stand for 5 min, centrifuge at 12000 r / min at 4°C for 15 min, carefully remove the supernatant, transfer to another centrifuge tube, add 100 ul of chloroform and isoamyl alcohol mixture (chloroform:isoamyl alcohol volume ratio 24:1), mix well and let stand for 5 min;

[0028] (2) Centrifuge at 4°C, 12000 r / min for 15 min, transfer the supernatant to another centrifuge tube, add 0.5 times the volume of the superna...

Embodiment 2

[0030] Example 2 Chicken articular cartilage RNA extraction

[0031] (1) Put 0.05 g of chicken articular cartilage stored at -80°C into a mortar filled with liquid nitrogen, grind it thoroughly until it becomes powdery, then add 0.5 ml of Trizol reagent (Invitrogen, Tankara, etc.), and wait for transformation When it is a solution, continue to grind thoroughly for 1 min; transfer the solution in the mortar to a 1 ml centrifuge tube (RNase-free, sterile), and mix vigorously. After standing for 5 min, add 100 ul of chloroform, shake vigorously, Let stand for 5 min, centrifuge at 4°C, 12000 r / min for 15 min, carefully remove the supernatant, transfer to another centrifuge tube, add 100 ul of chloroform and isoamyl alcohol mixture (chloroform: isoamyl alcohol volume ratio is 24 : 1), mix well and let stand for 5 min;

[0032] (2) Centrifuge at 4°C, 12000 r / min for 15 min, transfer the supernatant to another centrifuge tube, add 0.5 times the volume of the supernatant, 5 mol / L sod...

Embodiment 3

[0034] Example 3 Mouse Tibia Growth Plate Total RNA Extraction

[0035] (1) Put 0.05 g of mouse tibial growth plate stored at -80°C into a mortar filled with liquid nitrogen, grind it thoroughly until it becomes powdery, and then add 0.5 ml of Trizol reagent (Invitrogen, Tankara, etc.), Continue to grind thoroughly when it is converted into a solution, and the time is 1 min; transfer the solution in the mortar to a 1ml centrifuge tube (RNase-free, sterile), mix vigorously, let it stand for 5 minutes, add 100 ul of chloroform, shake vigorously to mix Evenly, let stand for 5 min, centrifuge at 12000 r / min at 4°C for 15 min, carefully remove the supernatant, transfer to another centrifuge tube, add 100 ul of chloroform and isoamyl alcohol mixture (chloroform:isoamyl alcohol volume ratio 24:1), mix well and let stand for 5 min;

[0036] (2) Centrifuge at 4°C, 12000 r / min for 15 min, transfer the supernatant to another centrifuge tube, add 0.5 times the volume of the supernatant, ...

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Abstract

The invention discloses a method for extracting RNA from animal cartilage tissue, relating to an RNA extracting method, for solving the problem that tibia has high content of polysaccharose substances, and thus the extraction of RNA is difficult. The method for extracting RNA from the animal cartilage tissue comprises the following steps: extracting RNA from the animal cartilage tissue by adoptinga Trizol reagent, adding mixed liquid of chloroform and isoamylol, taking the supernate, adding a sodium chloride solution and isopropanol, thus obtaining sediment RNA, carrying out dissolving by adopting DEPC treated water, adding diethyl ether, carrying out centrifugation, taking the lower clear liquid, adding diethyl ether, sodium chloride and isopropanol, carrying out ice-water bath precipitating, carrying out centrifugation, carrying out washing by adopting ethyl alcohol, carrying out dissolving by adopting DEPC treated water, and carrying out precipitating. The method provided by the invention is simple, is low in cost, short in consumed time, high in extraction number and quality, and wide in application, is suitable for RNA extraction of a tibia growth plate, and also can be applied to extraction of RNA of the cartilage tissue or bone tissue of the other parts.

Description

technical field [0001] The invention relates to an RNA extraction method, in particular to a method for extracting RNA from animal cartilage tissue. Background technique [0002] The tibial growth plate has more matrix and fewer cells. The matrix contains a lot of collagen and a lot of mucopolysaccharides. Among them, type II collagen is rich in hydroxylysine, and the hydroxyl groups of hydroxylysine are almost all combined with sugar, so the glycation rate High, so a considerable part of the tibial growth plate matrix is ​​polysaccharides. Since the physical and chemical properties of polysaccharides are very similar to ribonucleic acid RNA, when RNA is precipitated, proteoglycans and RNA are precipitated indiscriminately, resulting in polysaccharide-rich Gel-like precipitate, this kind of RNA precipitation containing polysaccharides is difficult to dissolve in water, or produces a viscous solution after dissolution, such RNA cannot be used for subsequent various molecular ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 田文霞陈书明宁官保高文伟张鼎宋涛张晨亮喻进
Owner SHANXI AGRI UNIV
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