Primers for identifying antelope horn and application thereof

A technology of primer pairs and antelope horns, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of unfavorable, cumbersome fluorescent staining methods, and long testing time

Active Publication Date: 2015-06-24
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In summary, different researchers have used protein electrophoresis methods to identify medicinal materials such as antelope horn, but these methods take a long time to test and r

Method used

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  • Primers for identifying antelope horn and application thereof
  • Primers for identifying antelope horn and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, the preparation and use method of the kit for identification antelope horn

[0051] 1. Design and synthesis of primer pairs for identification of antelope horns

[0052] RonM-t1:

[0053] 5'-TGTAAAAGGAGGGCCAGTGGMGCMCCMGATATRGCATTCCC-3';

[0054] VR1-t1:

[0055] 5'-CAGGAAACAGCTATGACTAGACTTCTGGGTGGCCAAA GAATCA-3'.

[0056] Select the universal primer COI (RonM-t1 / VR1-t) to amplify the samples of antelope horns, pair antelope horns and yellow sheep horns, gazelle horns, Przewalski horns, goose-throated gazelle horns, and goat horns, and then sequence the obtained sequences. For homologous comparison, design a pair of specific primers F1 and F2 according to the variable region, the sequence is as follows:

[0057] F1: 5'-ACTTCTAGCATTCTTCCATAGTTGAG-3' (SEQ ID NO: 1);

[0058] F2: 5'-GGGAAGTGAAAGGAGTAGGAGG-3' (SEQ ID NO: 2).

[0059] 2. Assembly of kits for identification of antelope horns

[0060] After the primers F1 and F2 designed and synthesized in ...

Embodiment 2

[0073] Embodiment 2, using the kit prepared in embodiment 1 to identify antelope horns

[0074] Samples to be tested: 6 samples shown in Table 1.

[0075] 1. Extract genomic DNA from the sample to be tested

[0076] Take the sample of the medicinal material to be tested, after surface disinfection with 75% (volume fraction) ethanol, use a sterilized scalpel to take 20 mg of the sample, then put it into a 1.5mL EP tube, grind it with a grinder, and use TIANDZ bone DNA Total DNA was extracted with an extraction kit (Beijing Tianenze Biotechnology Co., Ltd.) and stored at -20°C.

[0077] 2. PCR amplification

[0078] Using the genomic DNA extracted from the sample to be tested in Step 1 as a template, the primer pair (primer F1 and primer F2) designed in Step 1 of Example 1 was used for PCR amplification. The specific reaction system and reaction conditions are the same as Step 3 1 of Example 1. At the same time, double distilled water was used as a template in the experiment...

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Abstract

The invention discloses a primer pair for identifying antelope horn and application thereof. The primer pair for identifying or auxiliarily identifying antelope horn is particularly a primer pair composed of two single-chain DNA molecules disclosed as Sequence 1 and Sequence 2 in the sequence table. The experiment proves that the primers implement quick and accurate identification on antelope horn, mongolian gazelle horn, goa horn, Procapra przewalskii horn, goitered gazelle horn, goat horn and the like by quick PCR (polymerase chain reaction) and fluorescent dyeing. The fluorescent dyeing process is introduced to detect the identification result, thereby providing technical support for field application of drug molecule identification.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, relates to the field of identification of traditional Chinese medicines and medicinal materials, in particular to a pair of primers for identifying antelope horns and an application thereof. Background technique [0002] According to the first edition of "Chinese Pharmacopoeia" (2010), the source of antelope horns is the horns of the bovid saiga antelope Saiga tatarica Linnaeus. Antelope horn is a traditional precious Chinese medicinal material in my country. It has the effects of calming the liver and calming the wind, clearing the liver and improving eyesight, and dispersing blood and detoxification. In my country and some Central Asian countries, but the saiga antelope distributed in the Xinjiang region of my country has become extinct due to overhunting, and the population distributed abroad has also declined by 90% in just a few years due to poaching and other reasons (Jiang ZG, Gao...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 黄璐琦袁媛崔占虎蒋超金艳
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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