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Triplex PCR system detecting method of panda microsatellite site

A technology of microsatellite sites and detection methods, applied in the field of bioengineering, can solve problems such as not established, and achieve the effects of improving efficiency, saving DNA samples, and simple detection methods

Inactive Publication Date: 2017-09-22
四川省自然资源科学研究院 +1
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Problems solved by technology

Although microsatellite multiplex PCR technology has been widely used in other species (Wen Ping, Zhao Jian, Li Wei et al. Paternity identification of yellow-throated pseudowater turtle based on microsatellite multiplex PCR technology[J]. Acta Hydrobiology, 2015, ( 06):1134-1141.; Li Jiakai, Wang Zhiyong, Wei Xinjian, etc. Establishment and application of large yellow croaker microsatellite multiplex PCR system [J]. Fisheries Journal, 2014, (04):470-475.), but so far, Giant panda microsatellite multiplex PCR system has not yet been established

Method used

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  • Triplex PCR system detecting method of panda microsatellite site
  • Triplex PCR system detecting method of panda microsatellite site
  • Triplex PCR system detecting method of panda microsatellite site

Examples

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Embodiment 1

[0027] This example uses the giant panda samples collected at the Dujiangyan Giant Panda Base of the China Giant Panda Conservation and Research Center to establish a triple PCR system detection method. The specific method is as follows:

[0028] (1) DNA sample extraction: 14 blood samples were provided by the China Giant Panda Conservation and Research Center, and the giant panda blood genomic DNA was extracted according to the instructions of the Axygen blood genome extraction kit.

[0029] (2) Evaluate the size of the amplified product of the primer.

[0030] From the literature (Huang J, Li YZ, Du LM, et al. 2015.Genome-wide survey and analysis of microsatellites in giant panda (Ailuropodamelanoleuca), with a focus on the applications of a novel microsatellite marker system.BMC Genomics, 16: Article 61. ), find 15 pairs of giant panda microsatellite primers with good amplification effect, good typing effect and high polymorphism, Gpz6, Gpz47, Gpz20, Gpl47, Gpl29, Gpl60, Gpz54, G...

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Abstract

The invention belongs to the technical field of bioengineering, and particularly relates to a set of triplex PCR system detecting method of a panda microsatellite site. The set of triplex PCR system detecting method of the panda microsatellite site roughly includes the steps that 1, genome DNA is extracted; 2, the fragment sizes of amplification products of initial primers (a single pair of primers) are evaluated; 3, triplex PCR primer combinations are designed and optimized; 4, triplex PCR amplification is conducted, and conditions are optimized; 5, gene typing effects are detected, and a final combination mode is determined. The triplex PCR system detecting method of the panda microsatellite site is simple, compared with conventional panda microsatellite PCR systems, 66.7% of DNA samples and 66.7% of costs are saved, and the efficiency is improved by 200%. The triplex PCR system detecting method of the panda microsatellite site can be popularized in the aspects of evaluating panda paternity tests, genetic variation of populations and genetic structures.

Description

Technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to a set of detection methods of a triple PCR system for panda microsatellite sites. Background technique [0002] Microsatellite DNA is a short tandem repeat sequence in the genome of eukaryotes (Zietkiewicz E, Rafalski A, Labuda D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics, 1994, 20:176-183). Due to the large number of microsatellite sites, evenly distributed in the genome, co-dominant inheritance and easy detection, microsatellite DNA has been widely used (Wan QH, Wu H, Fujihara T, et al. Which genetic marker for which conservation genetics issue .Electrophoresis, 2004, 25: 2165-2176). Giant panda microsatellites have been used to solve paternity testing, genetic diversity, genetic structure and other issues (Li DS, Cui HM, Wang CD, et al. A fast and effective method to perform paternity testing ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2531/113C12Q2537/143C12Q2525/191
Inventor 朱英杨海琼李裕冬熊铁一
Owner 四川省自然资源科学研究院
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