9 results about "12s rrna" patented technology

The invention belongs to the technical field of molecular biology, and relates to a primer pair combination for identifying animal species and application of the primer pair combination in a kit. According to the method, the identification efficiency and accuracy are remarkably improved, PCR amplification is carried out by utilizing various gene markers (such as COI, cyt b, 16S rRNA and 12S rRNA), and individual differences of different families and even in the same species can be effectively distinguished due to high conservative property and variability of the gene regions in different species. For example, primer pair sequences SEQ ID NO.1-2 and 9-10 are designed for general animal species, and SEQ ID NO.3-4 are specially optimized for bird animal species, so that the design ensures that accurate results can be obtained even for complex sample types.

The invention discloses a species identification kit for animal mitochondrial bar code multiplex PCR amplification and nanopore sequencing, and relates to the technical field of species identification. The species identification kit comprises optimized primer pairs aiming at four hyper-variable genes (Cytb, COI, 16S rRNA and 12S rRNA) of mitochondria, by introducing R, Y, D and other degenerate bases at key sites, the binding capacity of the primers and template DNA of different species is remarkably improved, the problems that an existing universal primer is prone to mismatch, low in efficiency and even failed in amplification during cross-species amplification are solved, and the species identification kit has the advantages of being high in specificity, high in specificity and high in specificity. The method can be used for species identification of a mixed DNA sample containing at least two different animal species sources. The species identification primer combination is applied to preparation of an animal species identification kit, and the detection coverage range of the kit is expanded. According to the animal species identification method provided by the invention, a multiple PCR amplification system and a nanopore length reading sequencing technology are combined, congeneric sibling species with highly similar sequences can be distinguished, and the method has good accuracy.

The invention relates to the field of kits, in particular to a deafness-related gene mutation screening kit. The kit comprises an amplification primer, a PCR (Polymerase Chain Reaction) amplification enzyme reaction reagent, enzyme-free water, a magnetic bead purification reagent, a sequencing primer, a sequencing reaction reagent and a sequencing product purification reagent, wherein the amplification primer is designed for mutation of 26 sites in GJB2, GJB3, SLC26A4 and mitochondrial 12S rRNA genes; the magnetic bead purification reagent is used for purifying amplification products. An amplification primer follows the design principle that the distance between the amplification primer and a mutation site is at least 50 bases or above, and high-frequency single nucleotide polymorphic sites are avoided, PCR amplification adopts a falling annealing temperature control program, an amplification product is purified through a magnetic bead purification system, sequencing reaction configures an optimized working enzyme system, a specific thermal cycle program is adopted, and the high-frequency single nucleotide polymorphisms are obtained. And finally carrying out first-generation sequencing analysis. The method can realize high-specificity, high-sensitivity and high-stability detection of deafness-related gene multi-site mutation, and has the advantages of simplicity and convenience in operation, accurate result and wide application range.

The application provides a digital PCR kit for detecting highland invasive species red-eared slider and bullfrog, a set of red-eared slider specific primer probes, which are composed of a primer pair for a specific segment of a red-eared slider mitochondrial CYTB gene and a red-eared slider specific TaqMan probe matched therewith; a set of bullfrog specific primer probes, which are composed of a primer pair for a specific segment of a bullfrog mitochondrial 12S rRNA gene and a bullfrog specific TaqMan probe matched therewith, and a set of internal standard (IPC) primer pairs and internal standard TaqMan probes for monitoring the nucleic acid extraction and amplification reaction process monitoring. The application is provided with the primer pair for the specific segment of the red-eared slider mitochondrial CYTB gene and the red-eared slider specific TaqMan probe, the primer pair for the specific segment of the bullfrog mitochondrial 12S rRNA gene and the bullfrog specific TaqMan probe, and a common internal standard to construct a triple detection reaction system, thereby avoiding the quantitative uncertainty caused by the single species detection in the prior art which needs to be carried out in multiple reactions or relies on a standard curve for relative quantification.

The invention belongs to the technical field of biology, and discloses a DNA tetrahedral probe for live oocyte mitochondrial ATP imaging and application of the DNA tetrahedral probe. The invention provides a DNA tetrahedron delivery-based nucleic acid probe for fixed-point trigger cascade nucleic acid reaction, which can realize high-sensitivity and rapid detection of mitochondrial ATP of living oocytes. The nucleic acid probe is stable in structure and high in specificity, can accurately distinguish similar 12s rRNA and ATP analogues (such as CTP, GTP and UTP), remarkably reduces false positive signals, and ensures the reliability of detection results. The nucleic acid probe realizes ATP in-situ imaging at a mitochondrial level, is helpful for deep evaluation of an oocyte energy state, provides key indexes for quality reduction of oocytes of elderly women and evaluation of embryonic development potential, and has positive significance in improvement of assisted reproductive outcome.

This invention relates to the fields of molecular biology and forensic identification, specifically to a method for bird species identification based on multi-gene co-amplification and nanopore sequencing. The method involves extracting genomic DNA from avian biological samples; using the extracted DNA as a template, amplification is performed using four independent PCR primer pools that specifically target the avian mitochondrial cytochrome C oxidase subunit I gene, cytochrome B gene, 12S rRNA gene, and 16S rRNA gene; after purification of the amplification products, a nanopore sequencing library is constructed and sequenced; finally, bioinformatics analysis, including read length clustering, draft consensus sequence generation, and polishing, is performed on the raw sequencing data to obtain consensus sequences for each target gene. Species identification is then completed by comparing the sequence with a avian sequence database. This method effectively solves the problem that existing technologies cannot simultaneously meet the practical needs of large-scale, rapid, and high-success-rate identification of avian biological samples.

The application discloses a multiplex PCR specific identification primer for a pheasant meat source component, and the nucleotide sequence of a pheasant specific primer is 5'-GTAAAGTAGGACCATGCCTCT-3'. The application further discloses a kit containing the multiplex PCR specific identification primer, and a multiplex PCR identification method for the pheasant meat source component. The pheasant specific primer provided by the application can perform PCR amplification on 12S rRNA, the amplification effect is stable, a 151bp specific band can be generated, and the pheasant meat source can be effectively identified.

The invention discloses an animal species identification method based on a nanopore sequencing technology, and relates to the field of forensic genetics, the method comprises the following steps: (1) constructing a 12S rRNA gene reference database; (2) collecting a sample to be detected and extracting DNA; (3) carrying out PCR (Polymerase Chain Reaction) amplification by using a universal primer; (4) carrying out QNome nanopore sequencing; and (5) ClassIdent data analysis is carried out to confirm species. According to the animal species identification method based on the nanopore sequencing technology, a universal primer is adopted to simplify an experimental process, the long-reading-length portable nanopore sequencing technology is used to provide real-time sequencing data, and an automatic species identification process ClassIdent is constructed to perform data analysis, so that species identification can be performed on a sample with a single source, and the accuracy of species identification is improved. And the method has the characteristics of simplicity in operation, low cost, high accuracy and the like, and has a wide application prospect.

The invention discloses a DNA macro bar code primer group for identifying animal species in pet food, a kit and application of the DNA macro bar code primer group and the kit, and relates to the technical field of molecular biology. The primer group comprises two pairs of primers 12S rRNA and 16S rRNA, forward and reverse sequences of the primer group are respectively specific basic group arrangement, and the primer group can cover mammals, poultry and fish species. During application, pet food DNA (deoxyribonucleic acid) is extracted, PCR (polymerase chain reaction) amplification is carried out by the primer group (12S and 16S primers respectively adopt specific amplification procedures), after Sanger sequencing, comparison with an NCBI (National Center of Biotechnology Information) database is carried out, and species are determined by taking 98% sequence similarity as a threshold value. The primer group can deal with the DNA degradation problem caused by high processing, the detection sensitivity and accuracy are improved, and an effective technical means is provided for pet food quality control.