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Method for extracting and purifying RNA (Ribonucleic Acid) of rabbit vascular endothelial cell

A technology of vascular endothelium and rabbits, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of unsatisfactory, high cost, unsatisfactory results, etc.

Inactive Publication Date: 2013-12-18
HENAN UNIVERSITY
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Problems solved by technology

[0002] Rabbit arterial blood vessels are larger in diameter than mouse blood vessels, but it is difficult to obtain sufficient endothelial cell RNA
The results of the enzymatic hydrolysis and scraping methods reported in the literature are not ideal, it is difficult to avoid the contamination of smooth muscle cells, and the amount of RNA is very low, which cannot meet the PCR quantitative detection of 50 genes. If you want to obtain enough high-purity RNA samples, Many rabbits are needed to meet the needs, and the cost is relatively large (each purebred rabbit needs about 200 US dollars in the United States, and it takes about 30-50 breeds of rabbits to complete the quantitative PCR assay of 50 genes in 7 blood vessels, and the cost is 8,000 US dollars about

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  • Method for extracting and purifying RNA (Ribonucleic Acid) of rabbit vascular endothelial cell
  • Method for extracting and purifying RNA (Ribonucleic Acid) of rabbit vascular endothelial cell
  • Method for extracting and purifying RNA (Ribonucleic Acid) of rabbit vascular endothelial cell

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Embodiment Construction

[0014] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0015] Firstly, seven different blood vessel specimens were obtained in rabbits: left anterior descending coronary artery, right coronary artery, outer ring of aortic arch, inner ring of aortic arch, thoracic aorta, abdominal aorta, and iliac artery root close to the branch of abdominal aorta.

[0016] *Left and right coronary artery: Divide the heart into two, the left half has the left anterior descending coronary artery (LAD), and the right half has the right coronary artery (RCA), cut horizontally below the opening of the artery, Facilitate the passage of effluent. Connect a 27# needle and a 1ml syringe to the top of an ultrafine tip (a tip for Western blot loading), and rinse the blood vessel with 1ml of cold PBS for 3 times. From the upper opening of the coronary artery, 500 μl of Trizol (Invitrogen) was added, and the effluent was qui...

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Abstract

The invention discloses a method for extracting and purifying the RNA (Ribonucleic Acid) of a rabbit vascular endothelial cell. The method comprises extracting the RNA from left anterior descending coronary artery, right coronary artery, aortic arch outer ring, aortic arch inner ring, thoracic aorta, abdominal aorta, and iliac aorta. The method disclosed by the invention is high in extraction rate, and the extracted RNA is high in purity and can meet the requirements for detecting dozens of genes.

Description

technical field [0001] The invention belongs to the field of biology, in particular to a method for extracting vascular endothelial cell RNA from rabbits. Background technique [0002] Although rabbit arterial blood vessels are larger in diameter than mouse blood vessels, it is difficult to obtain enough endothelial cell RNA. The results of the enzymatic hydrolysis and scraping methods reported in the literature are not ideal, it is difficult to avoid the contamination of smooth muscle cells, and the amount of RNA is very low, which cannot meet the PCR quantitative detection of 50 genes. If you want to obtain enough high-purity RNA samples, Many rabbits are needed to meet the needs, and the cost is relatively large (each purebred rabbit needs about 200 US dollars in the United States, and it takes about 30-50 breeds of rabbits to complete the quantitative PCR assay of 50 genes in 7 blood vessels, and the cost is 8,000 US dollars For this reason, how to extract the RNA of ra...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 姬新颖李涛王芳芳朱诗白万振朱玉楠刘瑞敏张军白慧玲刘广超马远方
Owner HENAN UNIVERSITY
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