50 results about "12s rrna" patented technology

The invention relates to a method for detecting 20 hot mutation sites of four deaf genes such as GJB2, SLC26A4, GJB3 and 12S rRNA in a clinical sample, designing amplimers and single base extension primers of different sites aiming at abnormal SNP gene sites of the four genes by a matrix-assistant laser desorption / ionization flight time mass spectrum detection method and then carrying out mass spectrometry of a specific SNP site genotype.

The invention relates to a kit for building a library for detecting hereditary deafness genes and its application. The kit includes an amplification primer set for amplifying hereditary deafness genes; Primers designed for the polymorphism of the gene and the mutation site of the 12s rRNA gene. The present invention adopts multiplex PCR technology, which can detect more than 20 mutation sites of deafness genes in a single reaction; in conjunction with the "double label" system, the DNA amplification product of each sample has two sets of independent label sequences, so The amplification products of all samples can be mixed and sequenced simultaneously to achieve high-throughput detection, thereby greatly reducing the detection cost of a single sample.

The invention relates to the technical field of chemical detection and discloses a method for conducting real-time fluorescence identification on an edible bird's nest product through PCR. The method comprises the following steps: 1, extracting DNA of the edible bird's nest product to be detected; 2, conducting real-time fluorescence PCR amplification on esculent swift constituents, wherein firstly, 12.5 microliters of 2*PCR Master Mix, 0.5-1 microliter of an upstream primer, 0.5-1 microliter of a downstream primer, 0.5-1 microliter of Taqman probes and 1-2 microliters of the edible bird's nest product to be detected with the concentration being 50 ng / microliter are added into a PCR reaction tube, supplementing double distilled water to be 25 microliters, and conducting uniform mixing; secondly, putting the PCR reaction tube into a fluorescent quantitative PCR instrument to complete PCR amplification; 3, analyzing the amplification result with real-time fluorescence PCR instrument analysis software. The main principle of the method is that an edible bird's nest is mainly from saliva of esculent swifts and contains DNA of the esculent swifts, and realness of the edible bird's nest is identified by detecting the 12S rRNA gene of the esculent swifts in the edible bird's nest.

The invention discloses a fluorescent detection kit for detecting four deafness predisposing genes simultaneously. The kit comprises reagents before amplification and reagents after amplification, wherein the reagents before amplification comprise a polymerase chain reaction (PCR) buffer solution, a reaction mixture of MgCl2 and deoxyribonucleoside triphosphates (DNTPs), Taq DNA polymerase, ultrapure water, and a primer mixture for amplifying loci of GJB2235delC, 12S rRNA 1555A>G mutation, 1494C>T mutation, IVS7-2A>G and Amelogenin at high specificity; and the reagents after amplification comprise a genotyping standard and an internal standard. Deafness gene loci of the GJB2235delC, 12S rRNA 1555A>G mutation, 1494C>T mutation, IVS7-2A>G and Amelogenin are simultaneously detected at high sensitivity and high specificity by combining a fluorescent labeling technology, a linolenic acid (LNA) nucleoside monomer doping-primer modification technology and a capillary electrophoresis technology for the first time, manpower and material resources and time are greatly saved, and pollution due to multi-step operation is prevented.

The invention provides a deafness pathogenic gene detection kit utilizing a time-of-flight mass spectrometry. The kit comprises a reagent for detecting at least following 26 mutation sites of 12S rRNA(ribosomal Ribonucleic Acid), GJB2 and SLC26A4 genes: 12S rRNAm.1494-C is greater than T, 12S rRNAm.1555Ais greater thanG, GJB2c.35delG, GJB2c.257C is greater than G, GJB2c.427C is greater than T, GJB2c.176del16, GJB2c.9G is greater than A, GJB2c.235delC, GJB2c.299-300delAT, SLC26A4IVS4+2T is greater than C, SLC26A4c.1673A is greater than T, SLC26A4c.1520delT, SLC26A4c.2027T is greater than A, SLC26A4c.1975G is greater than C, SLC26A4c.1226G is greater than A, SLC26A4c.1318A is greater than T, SLC26A4c.1229C is greater than T, SLC26A4c.281C is greater than T, SLC26A4c.2168A is greater than G,SLC26A4IVS7-2A is greater than G, SLC26A4c.1174A is greater than T, SLC26A4c.235C is greater than T, SLC26A4c.1340delA, SLC26A4_c.589G is greater than A, SLC26A4c.916-917insG and SLC26A4c.IVS15+5G isgreater than A. The kit provided by the invention has the advantages of more mutation detection sites, high throughput, simplicity in operation and short period, high accuracy, high stability and lowcost.

The present invention discloses a fluorescence detection kit for detecting deafness susceptibility gene 12S rRNA 1555A>G, comprising a pre-amplification reagent and a post-amplification reagent, wherein the reagent before amplification includes a PCR buffer solution, a reaction mixture of MgC12 and dNTPs, a Taq enzyme, an ultra pure water, and a primer mixture for high- specificity amplification of 12S rRNA 1555A>G and Amelogenin loci; the post-amplification reagent includes a genotyping standard and an internal standard. The detection kit, with the loci of 12S rRNA 1555A>G and Amelogenin as detection objects, can screen out individuals having mutation at the above loci through the amplification of the deafness susceptibility gene locus, detection by capillary electrophoresis, and comparison between the detection object and the genotyping standard. The detection kit is of great significance to detection of deafness susceptibility gene and greater significance to deafness gene screening of the neonate. The detection kit is the first in the field of deafness gene screening to comprehensively combine fluorescence labeling technology, LNA nucleoside monomer incorporation-primer modification technology and capillary electrophoresis technology to realize detection of the deafness susceptibility gene locus 12S rRNA 1555A>G with high sensitivity and specificity.

The DNA probes produced by molecular cloning and the characterization of specific gene region sequences is provided, these can be used as genetic markers for the genes such as Cytochrome b (cyt b); Mitochondrial control region (D-Loop); Inter Transcribed Spacers (ITS2) and Rhodopsin (ROD), 12S rRNA and 16S rRNA in mesopelagic lantern fishes which are found in the mesopelagic zones of the oceans where the photic regime is of dim light and associate themselves with the oxygen minimum layer, it also includes the recombinant DNA techniques for the preparation of specific gene probes and sequences of species specific primers of lantern fishes, novel gene probes and novel oligonucleotides for amplification of myctophid genes are disclosed.

Method for analyzing a phyletic lineage of scallop, which comprises the steps of sequencing a mitochondrial DNA of the scallop to determine a nucleotide base sequence of the mitochondrial DNA, wherein the nucleotide base sequence includes a non-coding region containing a nucleotide base substitution locus which shows a sequence polymorphism indicative of a particular lineage of the scallop. Mitochondrial DNA of the scallop is amplified by PCR, using a suitable primer set designed on the basis of nucleotide base sequences conserved among known shellfish mitochondrial 16S rRNA and 12S rRNA genes, followed by sequencing to determine a non-coding region therein. In such non-coding region, a nucleotide base substitution locus is located, whereby a particular lineage of the scallop is determined. Based on those steps, a Japanese scallop, Patinopecten yessoensis, is analyzed as to the nucleotide base sequence and lineage thereof.

The present invention relates to one pair of PCR primers named as G-dloop, and provides one pair of amplification primers capable of amplifying the high variation zone of rockfish mitochondrial genome in high efficiency and its design method. The amplification primers consist of two single strand oligonucleotides. Through logging-on Genebank to search vertebrate mitochondrion DNA cytochrome b gene and the high conservation area of 16S rRNA gene sequence and homologous comparison, one pair of amplification primers is obtained. Through long PCR amplification, the target segments of 32 varietiesof rockfish are obtained and sequenced. The obtained sequences are compared by means of using homologous comparison software Clustal X 1.83 to fine the conservation sequences in the cytochrome b5' ends and the 12S rRNA 3' ends of the mitochondrial genomes of the 32 varieties of rockfish, and the said amplification primers are designed based on the degeneracy principle.

Method for analyzing a phyletic lineage of scallop, which comprises the steps of sequencing a mitochondrial DNA of the scallop to determine a nucleotide base sequence of the mitochondrial DNA, wherein the nucleotide base sequence includes a non-coding region containing a nucleotide base substitution locus which shows a sequence polymorphism indicative of a particular lineage of the scallop. Mitochondrial DNA of the scallop is amplified by PCR, using a suitable primer set designed on the basis of nucleotide base sequences conserved among known shellfish mitochondrial 16S rRNA and 12S rRNA genes, followed by sequencing to determine a non-coding region therein. In such non-coding region, a nucleotide base substitution locus is located, whereby a particular lineage of the scallop is determined. Based on those steps, a Japanese scallop, Patinopecten yessoensis, is analyzed as to the nucleotide base sequence and lineage thereof.

The invention relates to a hereditable deafness susceptibility gene screen test kit. The kit uses mutation from C to T of 1,494 locus of a 12s rRNA gene, mutation from A to G of 1,555 locus of the 12s rRNA gene, mutation from A to G of IVS7(-2) locus of an SLC26A4 gene and C(+ / -) of 235 locus of a GJB2 gene as detecting objects, designs and optimizes a set of specific primers respectively againsteach locus to be tested according to the PCR technical principle of a tetra-primer amplification refractory mutation system, amplifies the whole set of the primers in the same reaction tube, and performs primary multiple PCR amplification and primary gel electrophoresis on the four reaction tubes to obtain gene types of four loci simultaneously.