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Turtle 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: 2013-05-01
JILIN LEIBO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, DNA barcoding technology requires at least two gene loci to be selected, and selecting one locus alone cannot avoid the variation caused by mutations

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] 1. Pretreatment of test samples

[0059] Test samples (two kinds, 1 is genuine turtle shell, 2 is flower tortoise, fake turtle shell, all provided and identified by China Institute for the Control of Pharmaceutical and Biological Products), 2g is taken in each part, rinsed with pre-treatment solution after scrubbing clean, 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.

[0060] 2. Mitochondrial DNA Extraction

[0061] (1) Sample lysis Add 5ml of lysate to the pretreated sample, put it in a water bath, put it in a wat...

Embodiment 2

[0080] Example 2 Identification of commercially available turtle shell medicinal materials

[0081] 1. Materials

[0082] 5 kinds of turtle shell samples on the market, 1 genuine turtle shell (provided and identified by Jilin Drug Control Institute)

[0083] 2. Method

[0084] 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, put it in a centrifuge tube, add a corresponding volume of decalcification solution to each part at a mass-volume ratio of 1:20, and put it in a water bath at 56 °C for 48 h, with a rotation 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.

[0085] 2.2 Mitochondrial DNA extraction

[0086] (1) Sample Lysis Add 5ml of...

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Abstract

The invention relates to the technical field of traditional Chinese medicine identification, in particular to a turtle shell DNA (deoxyribonucleic acid) detection kit and an identification method. The turtle shell DNA detection kit comprises a sample pretreatment liquid, a decalcification liquid, a mitochondrial DNA extraction system, a PCR (polymerase chain reaction) system and a result observation system. The turtle shell DNA identification method comprises the steps of pretreating a detection sample, extracting mitochondrial DNA, conducting PCR primer design and synthesis, establishing PCR, and judging a result. A judgment standard is that a turtle shell is authentic if 1000bp and 500bp bands appear simultaneously, and the turtle shell is fake if only one or no band appears. The established multi-PCR identification method has the advantages of simplicity, convenience, quickness, reliable detection result and the like, and can differentiate and identify specificities of the turtle shell and the fake turtle shell accurately and simultaneously.

Description

technical field [0001] The invention relates to identification technology of Chinese medicinal materials, in particular to a test kit for turtle shell DNA and an identification method. Background technique [0002] Turtle shell is the carapace of Trionyx sinensis Wiegamann, an animal of the family Turtle family. Its original animal is Chinese soft-shelled turtle, commonly known as turtle, soft-shelled turtle, tortoise, etc., belonging to the class Repitlia; the order Testudinata; the family Tironychidae; and the genus Peodiscus. It is distributed in other regions of my country except Tibet, Qinghai and Xinjiang, and is most common in the Yangtze River Basin and South China. my country's soft-shelled turtle breeding output ranks first in the world, but since the 1990s, a large number of smuggled soft-shelled turtles have poured into many countries. In addition, the introduction and re-planting of various farms are extremely irregular, and they do not pay attention to cultiva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李明成苑广信张丽华夏薇王冰梅
Owner JILIN LEIBO TECH
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