83 results about "Amelogenin" patented technology

The invention relates to a compound amplification system for simultaneously analyzing a plurality of STR (short tandem repeat) loca, which is characterized by being used for carrying out compound amplification on 20 locas: Amelogenin, vWA, D21S11, D18S51, D5S818, D7S820, D13S317, D16S539, FGA, D2S1338, D19S433, D6S1043, D12S391, D8S1179, D3S1358, CSF1PO, Penta D, Penta E, TH01 and TPOX. The invention also relates to a method and a kit for simultaneously analyzing DNA (deoxyribonucleic acid) samples and other applications of the loca.

The invention relates to a Y-STR locus fluorescent label multiplex amplification system and an application thereof, and belongs to the field of polymorphic marker in detected human genome. The multiplex amplification system can simultaneously amplify loca of DYS19, DYS389-1, DYS389-2, DYS390, DYS391, DYS392, DYS393, DYS438, DYS439, DYA7.2 and Amelogenin. In the meanwhile, through the design of specific primers and a unique locus combination fluorescent label mode, only three marks can be adopted to simultaneously amplify the above loca, and amplified products are all controlled between 100bp to 400bp with very high sensitivity. A kit designed according to the amplification system is simple and rapid to operate, has high sensitivity, is high efficient, saves templates, has high individual identification capability, and reaches the level of foreign commercial kits of the same type.

The invention relates to the technical field of biology, and relates to a human X-chromosome composite amplification system composed of 20 short serially-connected repetitive sequences and applications thereof. The composite amplification system comprises 20 pairs of primers and is capable of amplifying 19 STR sites: DXS6795, DXS6803, DXS6807, DXS9907, DXS7423, GATA172D05, DXS101, DXS9902, DXS7133, DXS9810, GATA31E08, DXS6800, DXS981, DXS10162, DXS6809, GATA165B12, DXS10079, DXS10135, and HPRTB, and one sex recognition site: Amelogenin. Five-color fluorescence labeling is adopted by the system, the size of amplification products is in a range of 85 to 450 bp, the operation is simple, and the individual recognition performance is high. The sites provided by the provided composite amplification system are highly compatible to the conventional commonly-used X-STR kits, moreover, sites with high polymorphism information are provided, and the provided composite amplification system can be applied to paternity identification related with X-chromosome, individual recognition, and auxiliary detection of X-chromosome sex-linked hereditary diseases.

Compositions, methods and kits are disclosed for use in simultaneously amplifying at least 20 specific STR loci of genomic nucleic acid in a single multiplex reaction, as are methods and materials for use in the analysis of the products of such reactions. Included in the present invention are materials and methods for the simultaneous amplification of 23 and 24 specific loci in a single multiplex reaction, comprising the 13 CODIS loci, the Amelogenin locus, an InDel and at least six to ten additional STR loci, including methods, kits and materials for the analysis of these loci.

The invention discloses a compound amplification kit for InDel genetic polymorphic sites of human euchromosome and Y chromosome and application thereof. The kit comprises 47 pairs of euchromosome InDel site loca, 2 pairs of Y chromosome InDel site loca and a pair of specific amplification primers of a sex determination gene. The kit can be used for human individual recognition, paternity identification and degraded detection material recognition. The kit comprises 49 InDel sites which are relatively balanced in types and one sex determination gene. By adopting a six-colored STR fluorescence detection system, the kit is higher than the disclosed legal medical InDel detection kit. The kit disclosed by the invention is suitable for detecting Chinese groups. An amplification system of 50 sites is short in amplification fragment, and the amplicon is controlled at 200bp, so that the system is suitable for InDel typing of high degraded detection material; the amplification system improves the detection numbers of sites in the degraded detection material; the two Y-InDel sites introduced into the system plays an auxiliary judging role on the sex determination gene Amelogenin.

The invention discloses a fluorescence labeling combination amplification reagent kit capable of synchronously amplifying autosome gene loca and Y chromosome STR gene loca of people and application of the fluorescence labeling combination amplification reagent kit. The reagent kit can simultaneously amplify 34 gene loca to the maximum in a single tube reaction in a combination mode, wherein 34 gene loca include 15 autosome gene loca, 18 Y chromosome STR gene loca and Amelogenin. The six-color fluorescence technology is adopted, a unique gene locus arrangement mode is matched, the 34 gene loca are divided into five groups, all the groups of STR gene loca are labeled by different fluorescence labels, and in addition, the reagent kit has very high detection material adaptability. By means of the reagent kit, the autosome gene loca and the Y chromosome STR gene loca are detected simultaneously, the amplification template amount can be reduced, the detection time can be shortened, and meanwhile the efficiency of excluding and determining criminal suspects can be improved.

Process for purifying a recombinant protein including one or a few procedural steps only. The process combines the step of lysis of the host cell, with the purification of the protein of interest, allowing for a rapid and much more efficient process of purification. The conditions used during the purification process are those of a high temperature and a low pH, allowing for thermostable and acid-resistant recombinant proteins to be isolated from a suspension. The invention also relates to purifying recombinant proteins which are fusion proteins, wherein one part of the protein may be selected from an enamel matrix protein, such as amelogenin.

The invention provides a method and system for individual recognition and paternity identification of an unknown sample. The method includes: extracting the DNA of the unknown sample; acquiring the typing result of 17 loci contained by the DNA, i.e. 14 autosome STR loci, 2 Y-chromosome loci, and the sex determination locus Amelogenin, with the STR loci being D6S1043, D21S11, D7S820, CSF1PO, D2S1338, D3S1358, D13S317, D8S1179, D16S539, Penta E, D5S818, vWA, D18S51 and FGA, the Y-STR locus being DYS448, and the Y-chromosome insertion / deletion site being M134; and carrying out individual recognition and paternity identification according to the genotypes of the 17 loci of the unknown sample. The invention also provides the system for individual recognition and paternity identification of the unknown sample. The scheme involved in the invention can realize simultaneous detection of autosome STR loci and Y-chromosome loci, and also can shorten the detection time, thus improving the efficiency of individual recognition.

The invention relates to a composite amplification method and kit for STR loci. The composite amplification method and kit are used for amplifying 22 STR loci and one sex locus, wherein the 22 STR loci are composed of vWA, D21S11, D18S51, D5S818, D7S820, D13S317, D16S539, FGA, D2S1338, D19S433, D6S1043, D12S391, D8S1179, D3S1358, D1S1656, CSF1PO, Penta D, Penta E, TH01, TPOX, D10S1248 and DYS391, and the sex locus is Amelogenin. Amplimers used in the composite amplification method and kit comprise sequences as shown in SEQ ID No. 1 to 46. The composite amplification method and kit provided by the invention can acquire information about the 22 STR loci with good polymorphism and high individual discrimination power and about the one sex locus through simultaneous amplification at a time; and a composite amplification system composed of the loci has good balance, high sensitivity and high specificity, is accurate in typing results and can totally meet demands of judicial expertise.

The invention provides a kit for realizing multiple amplification of 21 short tandem repeats by adopting degenerate primers, and belongs to a multiple amplification system for simultaneously analyzing a plurality of STR gene loci. 21 gene loci are amplified in a multiple amplification system simultaneously, and are divided into the following four groups: the first group: D19S433, TH01, D21S11, D18S51 and Penta E; the second group: D3S1358, D13S317, D7S820, D5S818, CSF1PO and Penta D; the third group: Amelogenin, vWA, D8S1179, TPOX, D6S1043 and D1S1656; and the fourth group: D16S539, D12S391, D2S1338 and FGA. Specific primers are respectively designed on flanks of a gene loci repetitive sequence, so that the balance of the peaks of the fragments inside the gene loci groups achieves 40% or above, the four gene loci groups are subjected to multiple amplification, and the concentration of the primer of each gene locus is regulated, so that the integral balance of the peaks of the gene loci achieves 30% or above. With the kit, a balanced and stable multiple amplification result is easily obtained, so that the kit belongs to a multiple amplification system with low cost and high efficiency.

The invention discloses a human autosomal STR polymorphic site compound amplification kit and application thereof. The kit contains specific amplimers for amplifying the following 25 genetic locus: D3S1358, D13S317, D7S820, D16S539, D16S539, D1S1656, PentaE, TPOX, TH01, D2S1338, CSF1PO, Penta D, D19S433, vWA, D21S11, D18S51, D6S1043, Amelogenin, D8S1179, D8S1179, D5S818, D12S391, FGA, D22S1045, Yindel, D2S441 and D10S1248. The kit disclosed by the invention has the advantages that on the premise that the cumulative identification ability of the genetic loci is enhanced, the sensitivity is improved, all the genetic 25 loci can be detected under the condition that the DNA template amount is below 50pg and 29 cycle amplification is carried out, so that a complete genotype can be obtained. According to the human autosomal STR polymorphic site compound amplification kit disclosed by the invention, the quantity of the genetic loci can better satisfy continuous expansion of database construction and detection requirements of the genetic loci are added; the kit is efficient and stable, so that the detection rate of a detection material in a field case is improved and the expansion time isshortened.

The invention discloses an STR kit for a micro-amount degradation sample and application of the STR kit. The kit comprises 17 pairs of specific amplification primers of STR gene loci, the 17 pairs of specific amplification primers are divided into 5 groups, and fluorescent labels with 6 colors are related. The STR kit has the advantages of rapidness, high sensitiveness and high inhibition resistance on detection of micro-amount degradation samples. A mini type amplification system of the 17 gene loci established by the invention is short in amplified fragment, and is suitable for micro-amount, highly degraded or high-inhibitor-content difficult samples; and during actual application, STR types of special biological samples such as skeletons, contact cast-off cells and blood samples in soil can be obtained successively. DYS391 gene loci are introduced in a system, and the auxiliary judgment effect on sex genes Amelogenin can be achieved; and the kit can rapidly detect the samples, and the whole PCR process does not exceed 50 minutes.

The invention provides a multiplex amplification kit of 18 short tandem repeats (STR), relating to a multiplex amplification system which simultaneously analyzes a plurality of STR loca. The multiplex amplification kit is characterized by carrying out multiplex amplification on the following 18 loca: Amelogenin, D6S477, GATA198B05, D15S659, D8S1332, D3S3045, D17S1290, D14S608, D2S441, D18S535, D13S325, D10S1435, D11S2368, D1S1656, D7S3048, D10S1248, D19S253 and D3S1358. The invention also relates to a method and kit for simultaneously analyzing DNA (deoxyribonucleic acid) samples and applications thereof.

By utilizing a Mini-Primer strategy targeting the target site duplication (TSD) sequence of retrotransposons, insertion and null allele (INNUL) markers, which include short interspersed nuclear elements (SINEs), long interspersed nuclear elements (LINEs), and composite SVA retrotransposons (SINE / VNTR / Alu, where VNTR represents “variable number of tandem repeats” and Alu represents a type of primate specific SINE that has reached a copy number in excess of one million in the human genome), can be effectively used as markers for human identification and bio-ancestry studies regardless of the size of the inserted element. The size of the amplicons for INNULs and the difference between allelic states can be reduced substantially such that these markers have utility for analyzing high and low quality human DNA samples. Multiplexes including either 15 or 20 retrotransposable element (RE) markers plus Amelogenin for single tube amplification of DNA in four color detection were successfully designed. The multiplexes provided power of discrimination suitable for forensic and paternity analyses.

The present invention provides methods to determine gender of a canine subject, that include contacting a nucleic acid sample from the canine subject with at least one probe or primer specific for canine amelogenin, and using the binding of the at least one probe or primer to detect a difference between the canine amelogenin gene on the Y chromosome and the canine amelogenin gene on the X chromosome, thereby determining gender of the canine subject. In certain aspects, gender of the canine subject is determined by contacting the nucleic acid sample with a primer pair that generate different sized amplification products depending on whether an X chromosome or a Y chromosome copy of the canine amelogenin gene is amplified. In certain aspects and embodiments disclosed herein, in addition to detecting binding of at least one probe or primer to a canine amelogenin gene, methods of the present invention include detecting binding of at least one probe or primer to a canine microsatellite locus, thus providing a genotyping and gender determination assay.

The invention discloses a fluorescence labeled multiplex amplification kit for simultaneously analyzing 24 loci of human genome DNA, and an application thereof. The 24 loci comprise D16S539, D6S477, D15S659, D10S1248, D1S1656, D21S2055, D4S2366, D2S441, D3S1744, D22-GATA198B05, D3S3045, D11S2368, D17S1290, D10S2325, D14S608, D8S1132, D7S3048, D5S2500, D7S1517, D18S535, D10S1435, D13S325, D19S253 and a sex locus Amelogenin. The 24 loci are divided into 4 groups, and fluorescence labels with 5 colors are involved; the above fluorescence labeled multiplex amplification system has high sensitivity, and can detect all the 24 loci when the DNA template amount is 0.12ng; and the fluorescence labeled multiplex amplification system is suitable for being used in blood filter paper and direct amplification of FTA card acquired samples.

The invention provides a multiplex amplification kit containing 33 loca of a human genome. The multiplex amplification kit contains 18 A-STR loca which are recommended by a DNA database of the Ministry of Public Security to be used and also contains 14 Y-STR loca which are low in mutation rate and high in polymorphism and the Amelogenin locus, simultaneous amplification and detection on the 33 loca through a single tube is achieved, and synchronous amplification and detection on autosomes and Y chromosomes are achieved in a single experiment. The kit can directly amplify blood stains and saliva stains which take filter paper and an FTA filter as carriers without needing the template extraction and purification process and also can be suitable for DNA samples extracted through different extraction methods. The kit can be used for rapid investigation of a case, can improve the detection efficiency and increase the case investigation speed and especially has the significant effect on male sample trace detection in a raping case and father-child paternity test detection. The kit is wide in application range, good in compatibility and completely compatible with an existing legal medical expert DNA detection system.

The invention discloses a kit for human genotyping by fluorescence labeling STR and a detection method thereof, belonging to the field of human gene analysis and detection.The kit comprises a) a PCR amplification system: primer, PCR buffer solution, TaqDNA polymerase, template DNA, dNPT and MgCL2, b) detection reagent: deionized formamide solution and a GS500LIZ interior label; the primer is the primer of the following gene loca: Amelogenin, D8S1132, D12S391, GATA198B05, D7S3048, D2S1772, D11S22368, D6S1043 and D13S325.The detection method is carried out in accordance with the following steps: extracting DNA, carrying out compound amplification on gene loca, detecting and determining amplification products.Jointly applied to single parent identification and difficult genetic relationship identification cases with the existing commercialized kits, the invention can enable identification result to be more scientific and act as the beneficial supplement of the existing STR kit.

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 method and system for carrying out individual identification and paternity testing on Chinese populations and individuals. The method comprises: obtaining DNA of a to-be-tested individual; obtaining genotyping results of an autosomal STR locus, a sex determination locus Amelogenin and a Y-STR locus DYS391 of the DNA; and carrying out individual identification and paternity testing according to the genotyping results. The invention also provides the system for carrying out individual identification and paternity testing on Chinese populations and individuals. According to the scheme of the invention, by comprehensively analyzing the genotyping results of the autosomal locus and the sex determination locus, a genotyping result of the to-be-tested individual can be effectively obtained, and then a strong technical support is provided for public security organs to carry out individual identification and paternity testing.

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 invention provides method and system for Y-STR typing of an individual man. The method comprises the following steps: acquiring the DNAs of the individual man, wherein acquired DNAs comprise genotypes of 25 gene loci including 24 Y-STR gene loci and a gender determination locus Amelogenin, and the 24 Y-STR gene loci are DYS460, DYS389I / II, DYS390, DYS392, DYS458, DYS437, DYS385a / b, GATA_H4, DYS522, DYS456, DYS391, DYS447, DYS438, DYS448, DYS393, DYS635, DYS439, DYS19, DYS444, DYS527a / b, DYS617; and acquiring the Y-STR typing results of the individual man according to the 25 gene loci of the individual man. The Y-STR typing results can be used for family checking, male paternity testing, detection of male components in a hybrid male and female sample, detection of male components in a mixed sample of a plurality of males and the like for the individual man.

The invention discloses a fluorescent labeling complex amplification kit for insertion-deletion (InDel) polymorphism detection, and application of the kit. The kit comprises thirty-four InDel polymorphic sites and a sex locus (amelogenin). The fluorescent labeling complex amplification kit can be used for detecting the thirty-four InDel polymorphic sites and the one sex locus (amelogenin) at the same time; the kit can be used for detecting and genotyping 456 unrelated individuals from different ethnic groups from all over the country, has a cumulative personal identification rate reaching 0.99999985, and has a cumulative non-parent exclusion rate of 0.9989; the results prove that the kit is accurate in genotyping results, good in repeatability, high in sensitivity and accurate in genotyping results; therefore, a new method is provided for genetic relationship identification, sibling identification, individual identification, medical diagnosis, and the like.

The invention belongs to the technical field of medicine, and discloses a protein loading method, specifically adding a sodium tripolyphosphate solution dropwise into a chitosan quaternary ammonium salt solution containing a protein to be loaded, and then centrifuging and filtering the obtained mixed solution That's it. The method of the invention does not need to use organic solvents, has mild conditions and simple operation, and is very suitable for encapsulating protein drugs. The present invention uses it as a carrier, respectively carrying bovine serum albumin and recombinant amelogenin. The particle size of the bovine serum albumin-loaded nanoparticles is between 205.48-278.68nm, and the final release amount can be as high as close to 100%. The final release of loaded recombinant amelogenin nanoparticles can reach up to 50%.

The invention relates to the application of a composition containing recombinant amelogenin and propylene glycol alginate (“PGA”) on to cut dentin tubules followed by installation of a definitive restorative in a single visit.

The invention discloses a fluorescence detection kit for detecting deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T. The kit comprises a reagent before amplification and a reagent after amplification, wherein the reagent before amplification comprises a reaction mixture of a PCR (polymerase chain reaction) buffer solution, MgCl2 and dNTPs (deoxyribonucleoside triphosphate), Taq enzyme, ultrapure water, and a primer mixture for high-specific amplification of 12SrRNA1494C>T and amelogenin locus; and the reagent after amplification comprises an allelic ladder and an internal standard. According to the invention, 12SrRNA1494C>T and amelogenin locus are used as detecting objects, so that the fluorescence detection kit has important meaning to detection of deafness predisposing genes, particularly screening of newborn deafness genes.

The invention relates to a multiplex amplification system for analyzing a plurality of STR (short tandem repeat) gene loci simultaneously. The system is characterized in that 25 gene loci are subjected to multiplex amplification and include: Amelogenin, vWA, D10S1248, D2S441, D21S11, D18S51, D5S818, D7S820, D13S317, D16S539, FGA, D2S1338, D22S1045, D1S1656, D19S433, D6S1043, D12S391, D8S1179, D3S1358, CSF1PO, Penta D, Penta E, TH01, TPOX and DYS391. The invention further relates to a method and a kit for analyzing DNA (deoxyribonucleic acid) samples and applications of the method and the kit.

Compositions, methods and kits are disclosed for use in simultaneously amplifying at least 20 specific STR loci of genomic nucleic acid in a single multiplex reaction, as are methods and materials for use in the analysis of the products of such reactions. Included in the present invention are materials and methods for the simultaneous amplification of 23 and 24 specific loci in a single multiplex reaction, comprising the 13 CODIS loci, the Amelogenin locus, an InDel and at least six to ten additional STR loci, including methods, kits and materials for the analysis of these loci.

The invention discloses a composite amplification kit for simultaneously detecting autosomal and Y chromosome STR loci. The kit at least comprises specific amplification primer pairs of 40 loci, and the sequences of the primer pairs include SEQ ID NO.1 to SEQ ID NO.78. The kit can simultaneously detect 20 Y chromosome STR loci, 19 autosomal STR loci and Amelogenin locus, not only is accurate in typing, but also shortens the detection time, and improves the efficiency of removing and determining criminal suspects.

Isolated active compound of a naturally occurring fraction of Enamel Matrix Derivatives (EMD), having at least one of each two N-terminal polypeptide fragments of amelogenin, which are at least 95% identical to an amino acid sequence as shown in SEQ. ID. No:1 and / or 2. The invention relates to the use of said isolated active compound of a fraction and / or of the fraction itself, and / or the at least one of each two polypeptide fragments for use as a medicament and / or for the manufacture of a pharmaceutical composition for a variety of different medical indications such as inducing and / or promoting cementogenesis, bone growth and / or binding between parts of living mineralised tissue, for bonding of a piece of living mineralised tissue to a bonding site on a piece of other living tissue, for endorsing binding between hard tissues, for inducing regeneration of dentin, and / or for filling a mineralized wound cavity and / or tissue defect following from a procedure and / or trauma.