148 results about "Coi gene" patented technology

The invention discloses a PCR special primer pair for identifying pig-cattle-sheep derived ingredients in livestock and poultry meat based on mitochondrion COI gene, an identification method and a reagent kit, and belongs to the technical field of food detection. According to difference sites of COI genes of 9 kinds of animals including pigs, cattle, sheep, rabbits, pigeons, quails, chickens, ducks and geese, specific primers of 3 kinds of animals including the pigs, the cattle and the sheep are designed and are shown in a sequence table SEQ ID NO. 1, a sequence table SEQ ID NO. 2, a sequence table SEQ ID NO. 3, a sequence table SEQ ID NO. 4, a sequence table SEQ ID NO. 5 and a sequence table SEQ ID NO. 6. The identification method comprises the following steps: extracting total DNA of species to be determined, and performing PCR amplification. The reagent kit comprises SEQ ID NO. (1-6). The primers of the PCR special primer pair disclosed by the invention perform specificity amplification only on objective fragments of respective species and cannot generate a reaction with other species; the method is high in sensitivity, simple, convenient and rapid, and the pig derived ingredients, the cattle derived ingredients and the sheep derived ingredients in the livestock and poultry meat can be effectively identified, so that the adulteration phenomena of beef and mutton are avoided. Besides, designed primers SEQ ID NO. (1-6) are matched with related reagents, so that the reagent kit can be made, and favorable industrialized production and application prospects can be obtained.

The invention discloses a direct PCR detection method of Bombyx mori nuclear polyhedrosis pathogeny BmNPV, comprising the following steps of: firstly, treating the blood of infected Bombyx mori by ethanol precipitation to remove a PCR reaction inhibitor; designing the PCR primer of a polyhedrin gene according to the genome analysis of the Bombyx mori nuclear polyhedrosis pathogeny BmNPV; carrying out PCR amplification on the polyhedrin gene, adding an anti-PCR reaction inhibitor BSA (Bull Serum Albumin) to a reaction system and simultaneously carrying out PCR reaction by the primer of a Bombyx mori chondriosome CO I gene to make positive control; respectively getting the blood of normal Bombyx mori and the inflected midguts of the Bombyx mori infected with white muscardine, green muscardine, a bacterial disease and a microsporidia disease and then carrying out PCR reaction specification analysis; and detecting the PCR reaction result by agar gel electrophoresis. The whole detection process has simple operation and can be completed only by 4 hours. The disease can be diagnosed at the early stage of BmNPV infection by adopting the direct PCR detection method to carry out periodical sampling detection so as to adopt a measure in time to prevent the disease from happening.

The invention relates to a high-throughput multi-LDR parting kit for detecting farmland ecological environment, comprising mitochondria COI gene universal primers of 16 kinds of species in the farmland, specific probe sequences of the mitochondria COI gene universal primers of the 16 kinds of species in the farmland, and a reagent for PCR (polymerase chain reaction) proliferation and LDR (light dependent resistor) detection programs, wherein the mitochondria COI gene universal primers have the upstream of 5'-GGTCAACAAATCATAAAGATATTGG-3' and the downstream of 5'-taaacttcagggtgaccaaaaaatca-3'. In the invention, the kit effectively proliferates all target species sequences for the COI gene universal primers, and ensures that each target sequence and a low-copy template can be detected; the specific probe of each target species further ensures the specificity so as to totally distinguish each target sequence to realize multi-parting fixed quantity, thus the change of a farmland predation structure is known and the farmland ecological environment is maintained.

The invention relates to an amplification primer of a tridacnidae shellfish mitochondria COI (cytochrome oxidase I) gene. The known 52 tridacnidae sequences in NCBI (National Center for Biotechnology Information) are utilized, two tridacnidae sequences obtained by amplifying a universal primer are jointly utilized, a CLUSTALW software is used for comparison analysis, a plurality of pairs of primers are designed by using a primer design software Primer Premier 5 in a determined conversation region, 43 tridacnidae individuals are repeatedly subjected to PCR (Polymerase Chain Reaction) experiments in a 10 mul system and are finally detected through agarose gel electrophoresis, and an efficient amplification primer of the tridacnidae shellfish mitochondria COI gene sequence is sieved. The efficient amplification primer is characterized in that CQF is 5'-GATATTGGAACCCTTTACTT-3'; and CQR is 5'-TTCAGGATTGTCCCAAGAATCA-3'. By using the tridacnidae shellfish mitochondria COI primer, the target fragments of different specifies of tridacnidae shellfishes can be efficiently amplified; and the tridacnidae shellfish mitochondria COI primer has important significance for identifying tridacnidae species and carrying out phylogenetic analysis by using a molecular method, and can provide reliable technical guidance for developing and protecting tridacnidae germplasm resources by using the molecular means.

The invention discloses a culicoides bellulus specific gene and a molecular identification method thereof. The culicoides bellulus specific gene is characterized in that a molecular biology method is adopted to obtain the COI gene sequence of culicoides, and the species of the culicoides is identified by comparing gene sequence similarity. The ulicoides bellulus molecular identification method is beneficial for accurately and fast identifying culicoides bellulus.

The invention discloses a COI gene based PCR-RFLP discrimination method of seventeen economic sea cucumbers of Stichopodidae. The method comprises the following steps: 1, extracting DNA of a Stichopodidae sea cucumber to be detected as a detected sample, using the DNA as a template, and using primers COIe-F:5'-ATAATGATAGGAGGRTTTGG-3', COIe-R:5-GCTCGTGTRTCTACRTCCAT-3'PCR' to amplify a CO I fragment, wherein A is A / G; and 2, carrying out enzyme digestion on the CO I fragment by using a restrictive endonuclease group DdeI / SfcI or BstNI / Sau3AI, carrying out electrophoresis on the above obtained enzyme digestion product, estimating the size of each of the electrophoretic bands of samples to be detected, contrasting the number and the size of the electrophoretic bands in an electrophoretogram and the estimated value of the size with the reference band spectra of the seventeen sea cucumbers, and finding the standard spectral bands coupled with the number and the size of the electrophoretic bands, wherein the types represented by the standard spectral bands are the type of the detected sample. The method has the advantages of wide application range, simple operation, and accurate and fast detection, is especially suitable for the identification of a large batch of commodity sea cucumbers, so the method is a scientific method for identifying the sea cucumbers of Stichopodidae, and provides technologic supports for the standardization of the commodity market of the sea cucumbers and the protection of the sea cucumber resources.

The invention relates to a method for identifying Frankliniella occidentalis species by utilizing PCR (Polymerase Chain Reaction)-RFLP (Restriction Fragment Length Polymorphism) technology, comprising the following steps: (1) extracting Frankliniella occidentalis genome DNA; (2) conducting PCR amplification to the mitochondrion COI gene of the Frankliniella occidentalis genome DNA by taking the Frankliniella occidentalis genome DNA as a template; (3) conducting enzyme cutting to the PCR product prepared in step (2) by adopting restriction enzyme EarI to obtain an enzyme cutting product; and (4) conducting agar gel electrophoresis analysis on the enzyme cutting production prepared in step (3). The restriction enzyme is the common restriction enzyme, therefore, the method provides a simple, convenient and stable enzyme cutting marker for screening two types of Frankliniella occidentalis, and simultaneously explores and establishes an identification technology for the two types of Frankliniella occidentalis, and lays a foundation for identification on the population dynamics of the two types of Frankliniella occidentalis and research on biology and invasion mechanisms.

The invention discloses a PCR (Polymerase Chain Reaction) amplification kit for detecting clonorchis sinensis metacercaria on the basis of plastosome COI genes and an amplification primer. The PCR amplification kit comprises PCR amplification reaction liquid, wherein the PCR amplification reaction liquid includes 10mmol / L of Tris-HCl, 50mmol / L of KCL, 1.5mmol / L of MgCl<2>, 0.8mmol / L of dNTPs, 0.4[mu]mol / L of positive primers, 0.4[mu]mol / L of reverse primers, 20ng / [mu]L of DNA (Deoxyribonucleic Acid) templates and 0.075U / [mu]L of Taq enzyme. The PCR amplification kit has the advantages of high sensitivity, high specificity, high detection speed and high detection accuracy, and an effective technical measure is provided for diagnosis, treatment and prevention of clonorchis sinensis infection.

The invention belongs to the technical field of molecular biology and discloses a method for detecting material sources of meat products on the basis of short-sequence high-throughput sequencing. The method comprises the following steps that 1, genome DNA of the meat products to be detected is extracted; 2, universal primer sequences shown as SEQ ID NO:1 and SEQ ID NO:2 are designed and synthesized; 3, PCR amplification is carried out with the genome DNA of the meat products to be detected as a template to purify PCR products; 4, the PCR products are quantified, samples of the same quantity are mixed, a standard library is constructed, and then high-throughput sequencing is carried out; 5, COI sequence information of the meat products to be detected is obtained according to the sequencing result, so that the material sources of the meat products to be detected are obtained. By the utilization of the method, the material composition and the sources of the meat products can be identified, species can be well distinguished due to the difference between base sequences in the COI gene sequence, and the method is high in accuracy for identifying the material sources of the meat products.

The invention discloses a molecular marker for identifying coilia nasus and C. brachygnathus, and application of the molecular marker. The molecular marker is positioned on a COI gene of a mitochondrion of the coilianasus or the C. brachygnathus, and is specifically positioned at the part of 236bp of the COI gene of the mitochondrion; base variation information shows that T is more than C; the variation site of the coilianasus is T, and the variation site of the C. brachygnathus is C. A method for identifying the coilianasus and the C. brachygnathus can be used to identify the coilianasus and the C. brachygnathus by detecting the molecular marker. The method overcomes the disadvantages of only using experiential observation to identify the coilianasus and the C. brachygnathus, and provides a rapid and reliable molecular biological identification technology for the accurate identification of the coilianasus and the C. brachygnathus.

The invention discloses a double PCR detection method for authenticity identification of cordyceps sinensis, and solves the problem that in the prior art, the identification of cordyceps sinensis is conducted only regarding fungi, and therefore the prior art is short of sufficient accuracy. According to the method, in the same PCR reaction system, ITS of cordyceps militaris and COI gene nucleic acid fragments which are hosts of cordyceps militaris are simultaneously detected, and an amplification primer group for identifying the COI gene nucleic acid fragments is COI-F: GGAAATCCHGGATCTTTAATT and COI-R: GATGCCCCMGARTGTGCAAT; an amplification primer group for identifying the ITS is GITS-F10': GTTGCCTCGGCGGGAC and CITS-R10'-2: CMTTTGCTTGCTTCTTGACTGAG. The double PCR detection method for the authenticity identification of cordyceps sinensis is convenient to operate, high in cordyceps sinensis identification specificity and accuracy, good in repeatability and suitable for popularization.

The invention screens a pair of primers for amplification of a tegillarca granosa mitochondria COI sequence. The method comprises the following steps of: downloading the tegillarca granosa mitochondria COI sequence existing in the National Center of Biotechnology Information (NCBI); comparing and analyzing the tegillarca granosa mitochondria COI sequence by using BioEdit software; after determining a conserved domain, designing a plurality of pairs of primers by using primer design software Primer Premier 5; performing PCR reaction between the DNA of individual tegillarca granosa and the plurality of pairs of synthesized primers in a reaction system respectively; and finally, performing agarose gel electrophoresis detection to screen out a pair of primers COINF and COINR. Tests prove that the pair of primers has the advantages of single stripe and over 90-percent amplification efficiency, can meet the requirements of study on the genetic diversity of the tegillarca granosa, and plays an important role in the protection and utilization of tegillarca granosa resources, diversity estimation, genetic breeding and the like.

The invention discloses a molecular biological method for rapidly identifying Hoolock leuconedys and Nomascus leucogenys and belongs to the field of molecular biology. Genomic DNA is separated and extracted from to-be-detected objects and serves as a template; PCR is performed on the basis of a P1 primer pair and a P2 primer pair, and fragments A and B are obtained; a COI (cytochrome C oxidase I) gene complete sequence is obtained from the fragments A and B by splicing and cutting; species identification is performed according to the nucleotide sequence feature variation sites, disclosed for the first time, of the Hoolock leuconedys and the Nomascus leucogenys. Rapid and accurate molecular identification of the Hoolock leuconedys and the Nomascus leucogenys is realized for the first time, and various defects of the traditional morphological identification method are overcome. Identification objects include venous blood, feces, hair or urine samples of animals, and the method has the characteristics of being accurate, rapid and convenient, and has important function and significance for maintaining an ecosystem and protecting biodiversity.