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Method for detecting saiga tatarica horn ingredients in mixture and primers used in same

A mixture, antelope horn technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of high subjectivity

Active Publication Date: 2012-04-25
BEIJING TONGRENTANG CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the identification of antelope horns mainly uses traditional morphological methods, which rely on the experience of the identification personnel and are highly subjective. Therefore, an accurate and effective identification method is needed.

Method used

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  • Method for detecting saiga tatarica horn ingredients in mixture and primers used in same
  • Method for detecting saiga tatarica horn ingredients in mixture and primers used in same
  • Method for detecting saiga tatarica horn ingredients in mixture and primers used in same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Detection of Antelope Horn Components in Lingyang Qingfei Pills (Lingyangjiao Fang)

[0059] 1. Extraction of total DNA from antelope horns

[0060] This extraction step can be performed according to conventional operations in the field. The specific operation of this embodiment is as follows: antelope horns are ground into powder by liquid nitrogen, 200 μg is taken and added to 1 ml of lysate B (10 mM Tris-HCl pH8.0, 0.1M EDTA pH8.0, 0.5% SDS, 200 μg / ml proteinase K, 0.4M DTT), digested at 55°C overnight, digested samples were boiled at 100°C for 10min, ice-bathed for 5min, centrifuged at 12000rpm for 15min, and the supernatant was extracted with phenolform. Then precipitate the DNA with 2 times the volume of absolute ethanol, wash the precipitate twice with 75% ethanol, and dissolve in an appropriate amount of double distilled water. The sample DNA was purified by a PCR purification kit (Kangwei Century PCR Product Purification Kit 50T, product number CW05...

Embodiment 2

[0092] Example 2 Identification of Antelope Horn Components in Niuhuang Qingxin Pills (Antelope Horn Prescription)

[0093] Select 3 batches (10 pills / batch) of Niuhuang Qingxin Pills (Antelope Horn Prescription) produced by Tongrentang—a total of twenty-nine medicines, of which the Antelope Horn component accounts for about 0.9% (mass percentage) as the sample to be tested, extract The total DNA was subjected to single-plex PCR amplification with the primer pairs of SEQ ID Nos.1 and 2, SEQ ID Nos.3 and 4, and SEQ ID Nos.5 and 6, respectively. The extraction process of the total DNA of the sample to be tested, the PCR amplification and the analysis process of the amplification result are the same as in Example 1.

[0094] See results Figure 5 (Take the experimental results of one sample as an example, the rest are the same), using the DNA extraction method of the present invention and the designed antelope species-specific primers can effectively amplify the target fragment ...

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Abstract

The invention provides a method for detecting saiga tatarica horn ingredients in a mixture and primers used in the same. The method mainly comprises the steps of extracting total DNA from the mixture to be detected; with the extracted total DNA as a template, utilizing specific primers to carry out PCR (polymerase chain reaction) amplification; and analyzing and judging the amplification results,wherein the specific primers are selected from one pair, two pairs or three pairs of such primers as SEQ ID Nos.1 and 2, SEQ ID Nos.3 and 4 and SEQ ID Nos.5 and 6. The invention further provides a kit which comprises the primers and is used for detecting saiga tatarica horn ingredients in the mixture, especially the crude medicines needing differentiating to judge whether the crude medicines are saiga tatarica horns or the traditional Chinese medicine preparations using saiga tatarica horns or substitutes. The primers have higher specificity and sensitivity.

Description

technical field [0001] The present invention relates to a method for detecting components of antelope horns in mixtures, especially medicinal materials, using molecular biology techniques, in particular to a method for detecting components of antelope horns in medicinal materials, or traditional Chinese medicine preparations using antelope horns or substitutes, and the method used in the method specific primers. Background technique [0002] Antelope horn is a traditional precious Chinese medicine in my country, and it is the horn of the bovid animal Saiga antelope (Saiga tatarica Linnaeus). It has the effects of calming the liver and calming the wind, clearing the liver and improving eyesight, dissipating blood and detoxifying, and is used for the treatment of internal liver wind, convulsions, convulsions due to high fever, headache and dizziness. However, at present, the identification of antelope horns mainly uses traditional morphological methods, which rely on the expe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 赵兴波彭鹏秦晨范国强邱落胡爱平
Owner BEIJING TONGRENTANG CO LTD
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