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Method for realizing RNA (ribonucleic acid) overexpression by ovarian orthotopic injection and application of method

An in situ injection and overexpression technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of not finding the same or similar reports, save experimental costs, and improve expression levels. High, overexpressed good effect

Inactive Publication Date: 2016-07-20
SOUTH CHINA AGRI UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0002] So far no report identical or similar to the method of the present invention has been found

Method used

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  • Method for realizing RNA (ribonucleic acid) overexpression by ovarian orthotopic injection and application of method
  • Method for realizing RNA (ribonucleic acid) overexpression by ovarian orthotopic injection and application of method
  • Method for realizing RNA (ribonucleic acid) overexpression by ovarian orthotopic injection and application of method

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Embodiment 1

[0041] 1).PCR primer design. PCR primers were designed for the cDNA sequence of mouse ovary lncRNA-NONMMUT047274. The primer sequences are as follows:

[0042]

[0043] 2). PCR template DNA preparation. Total RNA was extracted from the ovarian tissues of C57BL / 6 mice and cDNA templates for PCR were obtained by reverse transcription. Use TaKaRa HSDNAPolymerase enzyme amplification. After the PCR reaction, take 3 μl of the amplification product and add 1 μl of 5×DNA loading buffer to electrophoresis in 1% agarose gel at 120V, observe whether there are bright bands under ultraviolet light, and prepare for the purification of the PCR product.

[0044] 3). PCR amplification product purification. Use TaKaRaMiniBESTAgaroseGelDNAExtractionKitVer.4.0 Gel Extraction Kit for purification of PCR amplification products.

[0045] 4). PCR amplification product and MOUSEZP3PROMOTER (JD#445 backbone vector) backbone vector enzyme digestion reaction. Use NEBXhoI and SmaI restriction e...

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Abstract

The invention belongs to the technical field of biology and discloses a method for realizing RNA (ribonucleic acid) overexpression by ovarian orthotopic injection and application of the method.RNA overexpression in a mouse ovary is realized by means of in-vivo transfection so as to provide a technical support for further researches on functions of RNA in the ovary. The method has the advantages that the defect of lack of targeting property of caudal vein over-expressed genes is avoided; transfection reagent consumption is reduced by dozens of times as compared with that of caudal vein injection, so that experiment cost is saved; excellent overexpression effects are achieved, and RNA expression level is raised remarkably as compared with that of a blank control group. The method which is minimally invasive has no influences on survival of mice.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and application for realizing RNA overexpression by ovarian in situ injection. Background technique [0002] No report identical or similar to the method of the present invention has been found so far. Contents of the invention [0003] In order to overcome the shortcomings and deficiencies of the prior art, the primary purpose of the present invention is to provide a method for realizing RNA overexpression by ovarian in situ injection. The invention realizes the overexpression of RNA in the mouse ovary through the method of in vivo transfection, and provides technical support for further research on the function of RNA in the ovary. [0004] Another object of the present invention is to provide the application of the method for achieving RNA overexpression by orthotopic ovarian injection. [0005] The object of the present invention is achieved through the f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/66C12N15/113
CPCC12N15/113C12N15/66C12N15/85C12N2310/14C12N2800/107
Inventor 杨世华李加宇吴绮琪方志浩黄群山
Owner SOUTH CHINA AGRI UNIV
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