Method for detecting mouse original components by specific primer

Abstract

The invention belongs to the technical field of the detection of mouse original components, and particularly relates to a method for detecting the mouse original components by a specific primer. The craft process of method comprises the following five steps: specific primer design synthesis, DNA (Deoxyribonucleic Acid) extraction, reaction system establishment, PCR (Polymerase Chain Reaction) amplification and the analysis result evaluation of the mouse original components. The specific primer is designed and synthetized according to the mitochondrial cytochrome b gene sequence of a mouse in Genbank, wherein the base sequence of the forward primer F of the specific primer is 5'-GCCTTATAATTAATTGGAGGTA-3', and the base sequence of the downstream primer F of the specific primer is 5'-AGCTATATTATGTGCTTGAT-3'; on the basis of the mitochondrial cytochrome Cyt b gene sequence of the mouse, a real-time quantitative PCR method is used for detecting the mouse original components so as to bring good specificity and repeatability. When the real-time fluorescence quantitative PCR method carried out by the primer is used for detecting the mouse original components, operation is simple, short time is consumed, and specificity is high. Secondary pollution caused by product amplification when an amplification product is taken out to carry out electrophoresis identification in a traditional detection method is avoided, and a new approach is provided for the detection of the mouse original components.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of detection of animal-derived components, and in particular relates to a method for detecting mouse-derived components with specific primers. The specific primers are used to detect the mouse-derived components by means of real-time fluorescence quantitative PCR (polymerase chain reaction). [0003] Background technique: [0004] PCR (polymerase chain reaction) uses DNA to denature into a single strand at a high temperature of 95°C in vitro, and at a low temperature of about 60°C, primers and single strands are combined according to the principle of complementary base pairing, and then the temperature is adjusted to DNA polymerization The most suitable reaction temperature for enzymes is 72°C. DNA polymerase synthesizes complementary strands along the direction from phosphoric acid to five-carbon sugar (5'-3'). The PCR instrument based on polymerase is actually a temperature-controlled device that can...

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Problems solved by technology

[0004] PCR (polymerase chain reaction) uses DNA to denature into a single strand at a high temperature of 95°C in vitro, and at a low temperature of about 60°C, primers and single strands are combined according to the principle of complementary base pairing, and then the temperature is adjusted to DNA polymerization The most suitable reaction temperature for enzymes is 72°C. DNA polymerase synthesizes complementary strands along the direction from phosphoric acid to five-carbon sugar (5'-3'). The PCR instrument based on polymerase is actually a temperature-controlled device that can , Refolding temperature and extension temperature are well controlled; polymerase chain reaction is a molecular biology technique used to amplify specific DNA fragments, which can be regarded as special DNA replication in vitro, PCR The biggest feature of DNA is that it can greatly increase a small amount of DNA, so whether it is the remains of ancient organisms or historical figures in fossils, or the hair, skin or blood left by the murderer in a murder case decades ago, as long as a little bit can be separated DNA can be amplified by PCR for comparison; mouse meat pretending to be expensive beef and mutton is a bad method often used by some greedy merchants. The dining table has brought great social impact and caused great personal injury to users. This is because rats themselves carry a variety of germs and are the storage hosts and main sources of infection for various infectious disease pathogens. At the same time, rat meat The content of toxic and harmful substances such as lead and arsenic is much higher than that of other meats. Direct consumption will bring great harm to people's health and the risk of disease transmission, ranging from poisoning to death; at present, there is no source of rats. The national standard or industry standard for the detection of sexual components is difficult for ordinary consumers and even food and drug quality supervision departments to identify with the naked eye, and the traditional detection method is cumbersome and time-consuming

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