Fluorescent quantitative PCR (Polymerase Chain Reaction) qualitative detection method for yak-derived fiber composition

A fluorescent quantitative and fiber-based technology, applied in the field of bioengineering, can solve the problems of high cost, unclear scales, time-consuming and labor-intensive electron microscopy, etc., and achieve the effects of good sensitivity, overcoming unclear scales, and high cost

Inactive Publication Date: 2012-08-15
上海出入境检验检疫局工业品与原材料检测技术中心 +1
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Problems solved by technology

[0005] The present invention is a qualitative identification method of yak cashmere fibers based on DNA detection technology, which overcomes the disadvantages of unclear scales, time-consuming and labor-intensive inspection under the existing optical microscope; and overcomes the shortcomings of high cost and cumbersome sample preparation by electron microscopy

Method used

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  • Fluorescent quantitative PCR (Polymerase Chain Reaction) qualitative detection method for yak-derived fiber composition
  • Fluorescent quantitative PCR (Polymerase Chain Reaction) qualitative detection method for yak-derived fiber composition
  • Fluorescent quantitative PCR (Polymerase Chain Reaction) qualitative detection method for yak-derived fiber composition

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Embodiment 1

[0054] 1. Design of primers and probes for real-time fluorescent PCR amplification of yak components

[0055] The sequences of mitochondrial 12SrRNA genes of goats, sheep and yaks were compared, and appropriate sequences were selected for primer design. The size of the amplified product of yak-derived fiber component-specific primers was 65 bp.

[0056] The sequences of specific primers and probes are as follows:

[0057] YakF: 5'-AAGCATCTACACCCCAGTGAGAAT-3' (SEQ ID NO: 1)

[0058] YakR: 5'-GCTTGATGCCAGCTCCTCTT-3' (SEQ ID NO: 2)

[0059] Pyak: FAM-CCCTCTAGGTTGTTAAAAT-MGB (SEQ ID NO: 3)

[0060] At the same time, common primers shared by yaks, goats, and sheep were designed. The size of the amplified product of the universal primers is 88bp. The sequences of the universal primers and probes are as follows:

[0061] GsyF: 5'-GGGTTTACGACCTCGATGTTG-3' (SEQ ID NO: 4)

[0062] GsyR: 5'-ACGTAGGACTTTAATCGTTGAACAAA-3' (SEQ ID NO: 5)

[0063] Pgsy: FAM-ACCGCTATCAAAGGTT-MGB (SEQ ID N...

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Abstract

The invention discloses a fluorescent quantitative PCR (Polymerase Chain Reaction) qualitative detection method for yak-derived fiber composition. The fluorescent quantitative PCR qualitative detection method comprises the following steps of: performing PCR amplification with the DNA (Deoxyribonucleic Acid) of yakwool as a template, and detecting fluorescent signals of amplification products, wherein a reaction system for the PCR amplification contains a specific primer pair for amplification of yak-derived fiber composition and specific probes for the yak-derived fiber composition. The fluorescent quantitative PCR qualitative detection method can be used for easily and quickly detecting and identifying whether the yak-derived fiber composition exists in textiles or not; and due to the good specificity and excellent sensitivity of the designed specific primer pairs and specific probes, the method can be used for detecting yakwool with proportion being 1%(wt).

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to a qualitative detection method for yak-derived fiber components by fluorescent quantitative PCR. Background technique [0002] The yak is a mammal that can survive in the harsh environment of the plateau and high cold, and it is also the only member of the bovine family that has textile utilization value. Yaks are divided into wild yaks and domestic yaks. Wild yaks are also called wild yaks. The scientific name is Bos mutus (Poěphagrt mutrs). Yak grows in the alpine region at an altitude of 3,000 meters to 5,000 meters, and can withstand the severe cold of minus 30°C to 40°C. Under the thick and long hair layer of the yak, there are fine and fine hairs. possible. There are more than 14 million yaks in the world, most of which are distributed in the Qinghai-Tibet Plateau and its surrounding alpine regions above 3000m above sea level. my country is the country ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 唐敏峰费静杨娟董正廉王卫华张小明吴海珍叶江
Owner 上海出入境检验检疫局工业品与原材料检测技术中心
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