Specific microsatellite primer composition for identifying experimental minipig breeds and method for identifying experimental minipig breeds

A miniature pig and microsatellite technology, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, recombinant DNA technology, etc., can solve the problem that primers are not specific between varieties, time-consuming and laborious results, and limit the experimental use of minipig varieties. Identification and other issues to achieve the effect of optimizing test conditions and methods of use, high accuracy and reliability

Active Publication Date: 2022-06-21
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

However, because the mining of microsatellites in the pig genome is not comprehensive enough, we can only use the microsatellite sequences of other species for homology comparison analysis, and the corresponding primers designed are not specific among breeds, and the identification of breeds is time-consuming and laborious and the results are inaccurate , limiting the identification of experimental miniature pig breeds

Method used

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  • Specific microsatellite primer composition for identifying experimental minipig breeds and method for identifying experimental minipig breeds
  • Specific microsatellite primer composition for identifying experimental minipig breeds and method for identifying experimental minipig breeds
  • Specific microsatellite primer composition for identifying experimental minipig breeds and method for identifying experimental minipig breeds

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Experimental program
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Effect test

Embodiment 1

[0038] A total of 160 4 experimental miniature pig breeds (48 Wuzhishan pigs, 15 Bama pigs, 50 inbred Luchuan pigs and 47 Tibetan pigs) were selected, and each individual ear tissue sample was collected and placed in a In the EP tube of 75% ethanol, the phenol chloroform method was used to extract genomic DNA, and NanoDrop 2000 (Thermo Fisher Scientific Company, USA), 1% agarose gel electrophoresis, and gel imaging (Beijing Zhongliwan Biotechnology Co., Ltd., Newbio Gi-1) Detect DNA concentration, OD value and DNA quality, and perform the following operations on DNA with DNA concentration ≥ 50 ng / μL, OD260 / 280 between 1.6 and 1.8, and the image of agarose gel electrophoresis is a highlighted band .

[0039] Fluorescent primers and common primers were synthesized by Shanghai Paisenuo Biotechnology Co., Ltd. The nucleic acid sequences of primers are shown in Table 1, and the PCR reaction system is shown in Table 2. Among them, the DNA concentration is 50ng / μL (concentration afte...

Embodiment 2

[0065] Collect the ear tissues of 10 individual experimental miniature pigs, identify the 10 pigs as individuals to be tested, number the individuals as #1-#10, and record the breed information corresponding to each individual number (including: # 1-#3 are Wuzhishan pigs, #4 are Bama pigs, #5-#7 are inbred Luchuan pigs and #8-#10 are Tibetan pigs) for comparison with the identification results of microsatellites. Each individual ear tissue sample was collected and placed in an EP tube filled with 75% ethanol, and genomic DNA was extracted by the phenol-chloroform method. NanoDrop 2000 (Thermo Fisher Scientific, USA), 1% agarose gel electrophoresis, Gel imaging (Beijing Zhongliwan Biotechnology Co., Ltd., Newbio Gi-1) was used to detect DNA concentration, OD value and DNA quality. Electrophoretic imaging of DNA with highlighted bands was performed as follows.

[0066] Fluorescent primers and common primers were synthesized by Shanghai Paisenuo Biotechnology Co., Ltd. The nucle...

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Abstract

The invention discloses a specific microsatellite primer composition for identifying experimental minipig varieties and a method for identifying experimental minipig varieties. The primer composition includes 3 pairs of primers. The present invention is based on the microsatellite sequence in the genome of the miniature pig breed, and obtains the specific microsatellite sequence and primers among different breeds of miniature pigs for experiments through population test verification and screening, optimizes the test conditions and usage methods of the primers, and solves the problem of Identification of miniature pig breeds for experimental use.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a specific microsatellite primer composition for identifying experimental miniature pig breeds and a method for identifying experimental miniature pig breeds. Background technique [0002] Experimental miniature pigs are one of the most widely used non-rodent large experimental animals in biomedical research. They have irreplaceable advantages over other experimental animals, and as the most likely donor for xenotransplantation, they have become a research hotspot. Research on minipigs and development and utilization by the biomedical community of general concern. The varieties and strains of miniature pigs used in experiments are various. In my country, there are typical Wuzhishan pigs, Bama pigs, inbred Luchuan pigs, Tibetan pigs, Tibet minipigs, Banna miniature pigs, Jianbaixiang pigs, Fernma miniature pigs, and Wujin pigs. Pigs, cooperative pigs, etc. Howe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/156C12Q2600/124C12Q2563/107
Inventor 王洪洋谈永松涂尾龙张莺莺吴华莉曹建国
Owner SHANGHAI ACAD OF AGRI SCI
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