Detection kit for quickly identifying donkey skin, horse skin and mule skin

A detection kit and mule skin technology, applied in the field of molecular biology, can solve the problem that horses, donkeys and mules cannot be identified at the same time, and achieve the effects of eliminating the inhibition of reaction conditions, high yield and good purity

Active Publication Date: 2014-09-17
VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If it is a donkey mule, it also has the mitochondrial gene of the donkey, and if it is a horse mule, it contains the mitochondrial gene of the horse, so only using the mitochondrial cytochrome B gene cannot simultaneously identify horses, donkeys and mules

Method used

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  • Detection kit for quickly identifying donkey skin, horse skin and mule skin
  • Detection kit for quickly identifying donkey skin, horse skin and mule skin
  • Detection kit for quickly identifying donkey skin, horse skin and mule skin

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Embodiment Construction

[0055] 1. Specificity test

[0056] Using genomic DNA extracted from mutton, beef, duck, fox, chicken, mink, and pork, and genomic DNA extracted from horses, donkeys, and mules as templates, fluorescence quantification was carried out according to the above-mentioned optimized reaction system and reaction conditions. PCR, testing the specificity of primers and probes. The 20μl reaction system contains (Table 1):

[0057] Table 1:

[0058] Reagent name

concentration

Dosage (μL)

HS-Taq

5U / μL

0.2

Premix

10X

2

Primer Mix

2uM

2

ProbeMix

2uM

2

IACDNA

1pg / ul

2

DNATemplate

20ng / μL

2

double distilled water

10.8

total capacity

20

[0059] The results showed that horses, donkeys, horse mules, and donkey mules all had CT values, while the other samples were not amplified. The results are shown in Table 2. It can be seen that the primers a...

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Abstract

The invention discloses a detection kit for quickly identifying donkey skin, horse skin and mule skin, which is used for accurately identifying donkeys, horses, mules and jennets by creatively applying 16SrRNA genes of CKM (creatine kinase) karyogene (nDNA) and mitochondrial genome DNA (mtDNA) at the same time from genetic background of the horses, the donkeys and the mules. The kit adopts an molecular beacon probe (MB) method to carry out a quintuple multicolor fluorescence quantitative PCR (polymerase chain reaction) detection technology, and also can detect donkey, horse, mule and jennet components; and moreover, the kit is added with exogenous internal reference as an interior label, can detect and avoid false negative results produced by PCR inhibitor contained in a sample. The quintuple multicolor fluorescence quantitative PCR is used for detecting in the same tube without opening a cover, so that the detection kit is not easy to pollute, and is accurate and stable, simple to operate, extremely high in sensitivity, strong in specificity, and the like, and therefore, a novel way is explored for identifying the donkey skin, horse skin and mule skin.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for quickly identifying animal species, in particular to a method for identifying donkey hide, horse hide and mule hide using a molecular beacon probe (MB) fluorescence quantitative PCR detection method and kits. Background technique [0002] Donkey-hide gelatin is a jelly made from the dehaired skin of an equine animal donkey. It has high medical and health value. Therefore donkey-hide gelatin occupies an important position in the market as a kind of health food, and is deeply favored by consumers. However, with the continuous development of the rubber market, the tight supply of raw materials for boiling rubber and the rising price of raw materials, some lawbreakers are now driven by interests to use cheap hides such as horse hides, mule hides, cow hides or pig hides to pass off as donkey hides. There are more and more phenomena, which bring huge hidden dan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6888C12Q2600/178C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 张全芳步迅刘艳艳
Owner VEGETABLE RES INST OF SHANDONG ACADEMY OF AGRI SCI
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