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Traditional Chinese medicine pheretima DNA detection kit and identification method thereof

A kit, the technology of Dilong, applied in the field of Dilong DNA detection kit and its identification, can solve the problems of time-consuming and high cost of DNA sequencing, and achieve the effect of simple identification method and reliable detection results

Inactive Publication Date: 2019-01-04
临沂市科学技术合作与应用研究院
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Sequence comparison found that these four genes have high homology in other types of earthworms and even other organisms. Unless DNA sequencing is performed in use, it is difficult to determine whether they are medicinal earthworms or even earthworms, and DNA sequencing is costly and time-consuming. duration

Method used

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  • Traditional Chinese medicine pheretima DNA detection kit and identification method thereof
  • Traditional Chinese medicine pheretima DNA detection kit and identification method thereof

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Effect test

Embodiment 1

[0029] Embodiment 1: a kind of Guangdilong DNA detection kit, it comprises:

[0030] (a) Identification reaction system: 100 μL of total system, 10 μL of buffer, 5 μL of 10 mM dNTP mixture, 11 μL of 0.1 mM primer, 1 μL of TaqDNA polymerase, 1 μL of sample to be tested, and make up the system with deionized water.

[0031] (b) Positive control reaction system: 100 μL of the total system, 10 μL of buffer, 5 μL of 10 mM dNTP mixture, 1 μL of 0.1 mM primer 1, 1 μL of TaqDNA polymerase, and 1 μL of Guangdilong DNA. Complete the system with deionized water.

[0032] (c) Negative control reaction system: total system 100 μL, buffer 10 μL, 10 mM dNTP mixture 5 μL each, 0.1 mM primer 1 1 μL, TaqDNA polymerase 1 μL, complete the system with deionized water.

[0033] (d) Negative control reaction system: total system 100 μL, buffer 10 μL, 10 mM dNTP mixture 5 μL, 0.1 mM primer 1 1 μL, TaqDNA polymerase 1 μL, cattle, sheep, chicken, duck intestines 1:1:1:1 Mix 1 μL of DNA and make up the...

Embodiment 2

[0055] Embodiment 2: a kind of Guangdilong DNA detection kit, it comprises:

[0056] (a) Identification reaction system: total system 100 μL, buffer 10 μL, 10 mM dNTP mixture 15 μL each, 0.1 mM primer 1 or primer 2 3 μL, TaqDNA polymerase 5 μL, sample to be tested 5 μL, deionized water Complete the system.

[0057] (b) Positive control reaction system: 100 μL total system, 10 μL buffer, 15 μL each of 10 mM dNTP mixture, 3 μL 0.1 mM primer 2, 5 μL Taq DNA polymerase, 5 μL earthworm DNA, complete the system with deionized water.

[0058] (c) Negative control reaction system: total system 100 μL, buffer 10 μL, 10 mM dNTP mixture 15 μL each, 0.1 mM primer 2 3 μL, TaqDNA polymerase 5 μL, complete the system with deionized water.

[0059] (d) Negative control reaction system: total system 100 μL, buffer 10 μL, 10 mM dNTP mixture 15 μL each, 0.1 mM primer 2 3 μL, TaqDNA polymerase 5 μL, cattle, sheep, chicken, duck intestines 1:1:1:1 Mix 5 μL of DNA and make up the system with deio...

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Abstract

The invention relates to the technical field of traditional Chinese medicine detection, in particular to a pheretima detection kit and a preparation method thereof. The kit comprises a genome extraction system, a PCR system and a detection system. The method comprises the steps of sample pretreatment, genome extraction, PCR, result identification and the like. Earthworm sample pretreatment involves dry samples and wet samples. According to an earthworm DNA detection method includes the steps that earthworm genome specificity oligonucleotide primers are designed, the primers are artificially synthesized, a reaction program is determined, and results are judged. Whether earthworms exist in medicinal earthworm raw materials or not is accurately judged. The identification method is simple, convenient and quick to use and reliable in identification result.

Description

technical field [0001] The invention relates to the technical field of detection of traditional Chinese medicines, in particular to a DNA detection kit for earthworm and an identification method thereof. Background technique [0002] Dilong is an earthworm that can be used as medicine, and it is a commonly used animal traditional Chinese medicine. From the investigation of the drug source and product identification of Earthworm in my country, it was found that there are mainly 13 species and varieties of protozoa of Earthworm in China, which belong to the 4 genera of Megaterridae, Orthowormidae and Streptococcus. In addition to the species collected and recorded in the Pharmacopoeia, it also includes the Hubei ringworm, the Zhili ringworm, and the Baijing ringworm. Due to the large number of original animal species of medicinal Earthworm, there is still a lack of real and effective identification methods, resulting in problems such as mixed varieties of medicinal materials ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/158
Inventor 高倩倩王伟于跃张振伟张焕琴
Owner 临沂市科学技术合作与应用研究院
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