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PCR detection primers and detection method for musk-derived ingredients

A technology of derived components and detection methods, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of molecular biological detection methods that are rare and have no musk deer-derived components , to achieve the effect of high sensitivity

Active Publication Date: 2016-04-27
珠海出入境检验检疫局检验检疫技术中心
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Problems solved by technology

At present, the DNA research on musk deer and its products mainly focuses on the extraction of DNA, and the research on the DNA detection and identification technology (non-identification to species) of the total class components required by the musk deer and its product market is relatively rare. Pseudo-identification is also limited to simple microscope observation and comparison method, and there is no molecular biological detection method for identifying musk deer-derived components

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  • PCR detection primers and detection method for musk-derived ingredients
  • PCR detection primers and detection method for musk-derived ingredients
  • PCR detection primers and detection method for musk-derived ingredients

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Embodiment Construction

[0021] The present invention will be further described below in conjunction with embodiment, but not limited thereto.

[0022] 1 Materials and methods

[0023] 1.1 Sample

[0024] Two musk deer samples (skin sheets) were reserved for our laboratory. The PCR product was cloned and sequenced using the vertebrate universal primer (ZH2006-1, CNAS approved standard) in the standard method of our laboratory, and its species source has been determined. 16 other animal samples (bear, deer, ox, antelope, donkey, horse, yak, sheep, dog, mink, camel, buffalo, tiger, pig, fox, rabbit) were retained in the laboratory.

[0025] 1.2 Main reagents

[0026] The DNA extraction kit E.Z.N.A.TMTissueDNAKit is a product of OMEGA Company in the United States, TIANGENTaqPCR Mastermix is ​​a product of Tiangen Biochemical Technology (Beijing) Co., Ltd.; DNAMarkerDL2000 is a product of TaKaRa, purchased from Treasure Bioengineering (Dalian) Co., Ltd.

[0027] 1.3 Main instruments

[0028] The 9600 ...

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Abstract

The invention discloses PCR detection primers and a detection method for musk-derived ingredients. The sequences of the primer pair are as shown in F-primer: TGGAGCTTTAACTAACTAGCCT, and R-primer: TGGATCAATGAGCGATATTGTG. According to the invention, the pair of specific primers is designed in a highly conserved domain in accordance with musk mitochondrion DNA (mtDNA) sequences revealed in GenBank; experimental results show that the primers are relatively good in specificity, specific amplification can be conducted on musk samples while cannot be conducted on other animal samples (including deer animals); and the primers are relatively high in sensitivity and are capable of detecting 102 copies / [micron]L according to the minimal order of plasmid copy magnitude. The method is applicable to rapid identification of common musk-derived ingredients on the market.

Description

technical field [0001] The invention belongs to the field of molecular detection, and in particular relates to a PCR detection primer and a detection method for musk deer-derived components. Background technique [0002] According to the Chinese animal classification code (GB / T15628.1-2009), musk deer belongs to the genus Musk deer of Mammalia Artiodactyla, and is specifically divided into: forest musk deer, horse musk deer, black musk deer, Himalayan musk deer and original musk deer. Into the musk subfamily in the deer family. Musk musk is a specialty species in eastern Asia. The musk extracted from the gonads of male musk deer living in cold areas can stimulate the central nervous system, break blood, and remove blood stasis when taken internally, and relieve pain and reduce swelling when used externally. It is an extremely valuable traditional Chinese medicine and spice with high economic value. Since the middle of the 20th century, due to the sharp increase in the dema...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 邵建宏罗宝正薄清如赵福振沙才华廖秀云王小玉徐海聂
Owner 珠海出入境检验检疫局检验检疫技术中心
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