Primer pair and application thereof in identifying pheretima asperillum

A technology of ginseng and primer pairs, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.

Active Publication Date: 2020-03-27
MUDANJIANG YOUBO PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the current identification of P. annulus relies on personal experience, so there is an urgent need for a method that can quickly and accurately identify P.

Method used

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  • Primer pair and application thereof in identifying pheretima asperillum
  • Primer pair and application thereof in identifying pheretima asperillum
  • Primer pair and application thereof in identifying pheretima asperillum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0044] 1. Reagents

[0045]COⅠ general primers LCO1490 and HCO2198 and specific primers dl301F and dl551R were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.; SpeedSTAR TM HS DNA polymerase (TaKaRa, batch number RR070A); 10 × FastBuffer I (TaKaRa company); dNTP Mixture (i.e. dNTPs) (TaKaRa company); DEPC water (Biosharp); agarose (Lonza); Mao Chemical Reagent Factory); 6 × Loading Buffer (TaKaRa Company); 10000 × SYBR GreenI nucleic acid dye (Beijing Soleibao Technology Co., Ltd.); DuRed nucleic acid dye (Beijing Fanbo Biochemical Co., Ltd.); D2000 DNA Marker (Beijing Days Root Biochemical Technology Co., Ltd.); TAE (50×) (Hefei Zhihong Biotechnology Co., Ltd.); Tks Gflex DNA polymerase and 2×Gflex PCRBuffer (2×Gflex PCRBuffer contains dNTPs) (TaKaRa).

[0046] 2 Species identification of samples to be tested

[0047] 1. Collect 25 samples of earthworms from different origins (i.e. samples to be tested), as shown in Table 1.

[0048] Table 1

[0049]

[0050]...

Embodiment 3

[0069] Example Three Primer Sensitivity Investigation

[0070] The concentration of template DNA is an important factor in specific PCR amplification, and a low template concentration requires high primer sensitivity. In this embodiment, the sensitivity of the primer pair of the present invention was investigated by changing the amount of DNA template used in the reaction system for the sample 1 to be tested. The amount of DNA template was adjusted to 25ng, 5ng, 1ng, 0.2ng, 0.04ng respectively, and the primer pair and amplification system of the present invention were used to carry out PCR reaction. The PCR reaction conditions were the same as in Example 2. PCR amplification results see image 3 . image 3 The 1-5 swimming lanes in the middle correspond to template dosages of 25ng, 5ng, 1ng, 0.2ng, and 0.04ng respectively. As can be seen from the results, bands can be amplified when the template DNA dosage is 25-0.04ng, which proves that the present invention The primer pa...

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Abstract

The embodiment of the invention provides a primer pair, a method for identifying pheretima asperillum by using the primer pair, and a kit for identifying the pheretima asperillum. By adopting the primer pair and the method provided by the embodiment of the invention, the DNA fragment in the pheretima asperillum can be specifically amplified, so that the pheretima asperillum can be accurately identified.

Description

technical field [0001] The invention relates to the technical field of the identification of the hair worm, in particular to a pair of primers and its application in the identification of the hair worm. Background technique [0002] Earthworm is a commonly used animal medicine. According to the 2015 edition of "Chinese Pharmacopoeia", Earthworm is Amynthas aspergillum, Pheretimav-ulgaris Chen, Pheretimaguillelmi (Michaelsen) or Ctenophora spp. Dried body of Pheretimapectinifera Michaelsen. Shenhuan hairworm is mainly produced in Guangxi, also known as Guangdilong. Due to its fine processing and definite clinical curative effect, it is recognized as the best quality and the highest price among Dilong Chinese medicinal materials. There are many animal species as the base source of the traditional Chinese medicinal materials of Dilonga, and the medicinal materials of Dilonga are mixed in the market. In addition, the wild resources of P. chinensis are decreasing day by day, res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888Y02A50/30
Inventor 李振国田晓轩王文秀张环宇倪开岭郝明张孝晨张立强贾力夫王丹丹何子龙
Owner MUDANJIANG YOUBO PHARMA CO LTD
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