Kit capable of quickly detecting real properties of leather and detecting method of kit

A kit and leather technology, which is applied in the field of kits for rapid detection of the real properties of leather, can solve the problems of sensory identification methods that are difficult to identify leather types, damage consumers' health and interests, and are expensive, and achieve effective supervision of leather products Quality, good economic and social benefits, and high sensitivity

Active Publication Date: 2012-11-28
GUANGZHOU QUALITY SUPERVISION & TESTING INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the wide variety of natural leather and the high price, in order to maximize profits, some unscrupulous traders pass off artificial leather as natural leather, or low-grade leather as high-grade leather, which damages the health and interests of consumers and affects the social economy and the development of the leather industry. had an adverse effect on
[0004] At present, the identification of leather materials at home and abroad is still mainly based on the sensory identification method of touching and seeing. With the improvement of leather processing technology, in some aspects, sensory identification methods are difficult to identify leather types, and more scientific development is urgently needed. effective method

Method used

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  • Kit capable of quickly detecting real properties of leather and detecting method of kit
  • Kit capable of quickly detecting real properties of leather and detecting method of kit
  • Kit capable of quickly detecting real properties of leather and detecting method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Kit for Rapidly Detecting the True Attributes of Leather

[0041] Contains the following components (for 50 PCR reactions):

[0042] (1) DNA extraction buffer (100mL)

[0043] Weigh 2.00mg CTAB (cetyltrimethylammonium bromide), 1.21mg Tris.HCl (trishydroxymethylaminomethane hydrochloride), 0.58g EDTA 2 Put Na (disodium edetate) and 8.18g NaCl in a volumetric flask, dilute to 100mL with water, and mix well. After sterilizing at 120° C. for 30 minutes, 10.00 mg of protease K (purchased from Dalian Bao Biological Co., Ltd.) and 15.43 g of dithiothreitol (DTT) were added.

[0044] (2) Taq DNA polymerase (25μl)

[0045] Purchase 25 μl of Taq DNA polymerase (125 IU in total) (purchased from Dalian Bao Biology Co., Ltd.).

[0046] (3) PCR reaction solution (1000μl)

[0047] Firstly, primers were prepared, and the prepared primer concentration was 100 nmol / L.

[0048] Primers were prepared according to conventional methods in the art. Synthesis was carried out ...

Embodiment 2

[0064] Example 2 Using the kit of Example 1 to detect genetic components in leather samples

[0065] The detection method comprises the following steps:

[0066] (1) Preparation of sample DNA

[0067] a leather sample

[0068] Cut 50cm 2 The left and right leather samples soaked and sterilized by alcohol were put into a shredder and crushed. Use phosphate buffer solution to soak the crushed leather sample for 12 hours under constant shaking conditions, during which the buffer solution is changed every 2 hours to reduce the content of pigment and salt ions such as chromium ions in the leather sample. After soaking, break the leather sample again, take 3 mL of the broken sample, add 2 mL of DNA extraction buffer, and mix well. Place the centrifuge tube at 65°C for 45-60 minutes, and shake it from time to time to fully lyse the sample. Take it out, add 1mL saturated sodium chloride solution, shake vigorously for 3-5min, place at 0°C for 5-10min, and centrifuge. The supernat...

Embodiment 3

[0098] Embodiment 3 reliability experiment

[0099] For the detection of main varieties of leather samples such as cow leather, sheep leather, and pig leather, this example uses real-time fluorescent PCR detection to verify the reliability of the method in Example 2.

[0100] After the DNA of the leather sample was extracted, a real-time fluorescent PCR detection experiment was carried out on the leather DNA sample. Take 948.0 μl of PCR reaction solution, 6.0 μl of Taq DNA polymerase, and 6.0 μl of probe in a centrifuge tube, mix well, distribute 48.0 μl into eight-tube tubes, and add 2.0 μl containing the target gene to the positive control tube For fragmented DNA samples, add 2.0 μl of control sample DNA without the target gene to the negative control tube, add double distilled water to the blank control tube, and then add 2.0 μl of the leather sample DNA to be analyzed to the remaining reaction tubes (reaction The system is 50 μl), centrifuged and placed on a real-time flu...

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Abstract

The invention discloses a kit capable of quickly detecting real properties of leather, and a detecting method of the kit. The kit comprises deoxyribonucleic acid (DNA) extraction buffer liquid, Taq DNA polymerase, polymerase chain reaction (PCR) liquid and sampling buffer liquid. The kit can be used for analyzing and identifying the compositions of genes of a leather sample from a genetics perspective, overcomes the shortcomings of conventional sensory identification, has the advantages of strong specificity, high sensitivity, good method reproducibility, rapidness, simplicity and convenience in detection and the like, fills the blank of leather gene detecting methods at present, serves as an accurate and easy-to-standardize analyzing method for identification of leather types, provides technical support for quality supervision and the like, can be widely applied to gene extraction of samples of leather including fur and the like, leather type identification and the like, has good economic and social benefits and great popularization and application values, and has major significance on effective supervision of leather product quality and protection of benefits of vast consumers.

Description

technical field [0001] The invention relates to a kit and a method for rapidly detecting the true attributes of leather. Background technique [0002] PCR is the abbreviation of polymerase chain reaction, which is a technique widely used in molecular biology. The main principle is to achieve a large amount of amplification of target gene fragments through thermal cycling. At present, this technology is more and more widely used in product testing. For example, use PCR technology to detect plant components in milk or sausage, use PCR technology to detect animal-derived components in products such as meat and bone meal, use PCR technology to detect genetically modified components in food, etc. [0003] Leather is mainly processed from mammalian animal fur and is an important export product for my country. Natural leather is classified according to its animal origin, mainly including cow leather, sheep leather, pig leather, horse leather, rabbit leather and so on. Due to the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 罗海英柯振华吴玉銮郭新东刘春生冼燕萍陈筱婷陈意光罗东辉吴文海
Owner GUANGZHOU QUALITY SUPERVISION & TESTING INST
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