Primer and probe for real-time fluorescence PCR (polymerase chain reaction) detection of alpaca component

Abstract

The invention belongs to a qualitative detection technology of animal-derived ingredients in feather down products, and specifically relates to a primer / probe group for detecting an alpaca component in a fiber product. A group of specific primer and probe is designed by selecting a mitochondrial COXI gene sequence of alpaca. By using the primer and the probe and adopting a real-time fluorescence PCR (polymerase chain reaction) technology, the alpaca component in the fiber product can be fast, sensitively and specifically detected. The primer and the probe can be provided together with other reagents in the form of a kit and be used for nucleic acid amplification reaction. The method is simple and convenient to operate and good in repeatability.

Description

technical field

[0001] The invention relates to a method for rapid detection of animal-derived components using nucleic acid amplification technology, in particular to a primer and a probe for real-time fluorescent PCR detection of alpaca hair components. Background technique

[0002] The English name of the alpaca is Alpaca, which is a kind of camelid, whose ancestor is a pack horse. Alpaca is full of treasures, and the alpaca fiber it produces is one of the world's famous high-grade animal fibers, known for its luxurious feel and luster. Because alpaca fiber contains a medullary cavity, its thermal insulation is better than other fibers, and it has excellent properties such as wrinkle resistance, abrasion resistance, and moisture retention. Fashions made of it are soft and warm, comfortable to wear, good in drapability, bright in color, and also emit colorful light under sunlight, which is very popular among consumers and has become another high-end special animal fiber p...

Examples

Embodiment 1

[0039] 1. Take samples from unused parts of fiber products, and take a total of about 1g of samples; cut the samples with scissors, break up and mix them with an analysis grinder, then use scissors to try to cut them to less than 2mm, and then use an analysis grinder to mix them evenly ;

[0040] 2. Weigh about 5 mg sample from the mixed sample, put it into a 2mL centrifuge tube, extract 2 tubes in parallel for each sample, add 180 μL of buffer GL (Buffer GL), 20 μL of proteinase K (Proteinase K) and 10 μL of ribonuclease A (RNase A, 10 mg / mL), warm in a 56°C water bath until the tissue is completely lysed for 3 hours; if the remaining fibrous tissue cannot be completely lysed, centrifuge at 12,000 rpm for 2 minutes after lysing, and then remove impurities. Follow up. Add 200 μL of Buffer GB (Buffer GB) and 200 μL of 100% ethanol to the lysate, and mix well by pipetting. During the warm bath, the sample can be taken out from time to time for shaking or pipetting to speed up ...

technical field

[0001] The invention relates to a method for rapid detection of animal-derived components using nucleic acid amplification technology, in particular to a primer and a probe for real-time fluorescent PCR detection of alpaca hair components. Background technique

[0002] The English name of the alpaca is Alpaca, which is a kind of camelid, whose ancestor is a pack horse. Alpaca is full of treasures, and the alpaca fiber it produces is one of the world's famous high-grade animal fibers, known for its luxurious feel and luster. Because alpaca fiber contains a medullary cavity, its thermal insulation is better than other fibers, and it has excellent properties such as wrinkle resistance, abrasion resistance, and moisture retention. Fashions made of it are soft and warm, comfortable to wear, good in drapability, bright in color, and also emit colorful light under sunlight, which is very popular among consumers and has become another high-end special animal fiber p...

Examples

Embodiment 1

[0039] 1. Take samples from unused parts of fiber products, and take a total of about 1g of samples; cut the samples with scissors, break up and mix them with an analysis grinder, then use scissors to try to cut them to less than 2mm, and then use an analysis grinder to mix them evenly ;

[0040] 2. Weigh about 5 mg sample from the mixed sample, put it into a 2mL centrifuge tube, extract 2 tubes in parallel for each sample, add 180 μL of buffer GL (Buffer GL), 20 μL of proteinase K (Proteinase K) and 10 μL of ribonuclease A (RNase A, 10 mg / mL), warm in a 56°C water bath until the tissue is completely lysed for 3 hours; if the remaining fibrous tissue cannot be completely lysed, centrifuge at 12,000 rpm for 2 minutes after lysing, and then remove impurities. Follow up. Add 200 μL of Buffer GB (Buffer GB) and 200 μL of 100% ethanol to the lysate, and mix well by pipetting. During the warm bath, the sample can be taken out from time to time for shaking or pipetting to speed up ...

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