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Tortoise shell DNA detection kit and identification method

A technology for detection kits and identification methods, applied in the field of identification of traditional Chinese medicinal materials, to achieve reliable detection results

Inactive Publication Date: 2013-08-21
JILIN LEIBO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, selecting a locus alone cannot avoid mutation-induced variation

Method used

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  • Tortoise shell DNA detection kit and identification method
  • Tortoise shell DNA detection kit and identification method
  • Tortoise shell DNA detection kit and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] 1. Pretreatment of test samples

[0064] Test samples (four kinds, 1 is authentic tortoise shell, 2 is tortoise shell with concave shell; 3 is tortoise shell of yellow-fronted box turtle; 4 is tortoise shell of Malayan box turtle, all of which are counterfeit products, provided by China Institute for the Control of Pharmaceutical and Biological Products and identified), take 2g for each portion, rinse with pretreatment solution after scrubbing, dry at room temperature, and irradiate with ultraviolet light for more than 30 minutes. Use a mortar to grind the sample to about 2-3 mm, place it in a centrifuge tube, add a corresponding volume of decalcification solution to each portion at a mass-volume ratio of 1:20, and bathe in a water bath at 56°C for 48 h at a speed of 100 r / min. Replace with fresh decalcification solution every 12 hours. After decalcification, centrifuge at 8000r / min for 5min, discard the supernatant, and precipitate for use.

[0065] 2. Mitochondrial ...

Embodiment 2

[0087] Example 2 Identification of commercially available tortoise shell medicinal materials

[0088] 1. Materials: 6 commercially available tortoise shell samples (labeled 4-9). One sample of turtle shell (label 10), one genuine turtle shell (both provided and identified by Jilin Drug Control Institute).

[0089] 2. Method

[0090] 2.1 Pre-treatment of test samples Take 2g of each test sample, rinse with pre-treatment solution after scrubbing, dry at room temperature, and irradiate with ultraviolet light for more than 30 minutes. Use a mortar to grind the sample to about 2-3 mm, place it in a centrifuge tube, add a corresponding volume of decalcification solution to each portion at a mass-volume ratio of 1:20, and bathe in a water bath at 56°C for 48 h at a speed of 100 r / min. Replace with fresh decalcification solution every 12 hours. After decalcification, centrifuge at 8000r / min for 5min, discard the supernatant, and precipitate for use.

[0091] 2.2 Mitochondrial DNA ...

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Abstract

The invention relates to the technical field of the identification of a Chinese medicine, and specifically relates to a tortoise shell DNA detection kit and identification method. The tortoise shell DNA detection kit comprises the following four parts: sample pretreatment fluid and decalcifying solution, a mitochondria DNA extraction system, a PCR reaction system and a result observing system. The tortoise shell DNA identification method comprises the following five steps: detection sample pretreatment, mitochondria DNA extraction, PCR primer design and synthesis, PCR reaction establishment and result determination. The determination standards are that if 100bp and 400bp strips simultaneously appear, the tortoise shell is an authentic tortoise shell, and if only one or no one appears, the tortoise shell is a fake tortoise shell. The multiple identification method provided by the invention has simple, quick, and reliable in the detection results and can accurately and simultaneously identify the specificities of the authentic tortoise shell and the fake tortoise shell.

Description

technical field [0001] The invention relates to identification technology of Chinese medicinal materials, in particular to a test kit for tortoise shell DNA and an identification method. Background technique [0002] The tortoise shell is the dry plastron and carapace of the tortoise chinemysreevesii (Gray) of the turtle family. The tortoise shell contains animal glue, keratin, fat, collagen, amino acid, and various constant and trace elements such as calcium, phosphorus, strontium, zinc, copper, etc., and is flat in nature. It has the effects of nourishing yin and suppressing yang, nourishing kidney and strengthening bones, nourishing blood and invigorating the heart; it is used for hot flashes due to yin deficiency, night sweats caused by bone steaming, false yellow blood, insomnia and forgetfulness. It is a commonly used precious medicinal material. Its medicinal materials are mainly produced in Hubei, Anhui, Hunan, Jiangsu, Zhejiang and other places. In recent years, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李明成苑广信张丽华夏薇王冰梅傅桂莲
Owner JILIN LEIBO TECH
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