201 results about "Str loci" 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.

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 relates to a composite amplification system for simultaneously analyzing multiple Y-STR loci. The composite amplification kit is characterized by compositely amplifying the following 26 Y chromosome loci: DYS19, DYS388, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385a, DYS385b, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, DYS460, DYS481, DYS533, DYS549, DYS570, DYS576, DYS643, DYS449 and Y-GATA-H4. The invention also relates to a method and a kit for simultaneously analyzing DNA samples, and applications thereof.

The invention discloses a fluorescent labeling composite amplification kit for 30 STR (short tandem repeat) loci of human Y chromosome and application of the kit. It is possible to multiple amplify 30 low-mutation-rate STR loci on Y chromosome in single reaction; by designing locus arrangements and specific primer sequences, 30 loci are divided into 5 groups, and six-color fluorescent system marks are used; the Y-STR kit is the domestically first one employing six-color fluorescent detection technology and is an STR kit detecting a highest number of loci, all the loci included are low-mutation-rate Y-STR loci, and the kit is more suitable for examining male families; the composite amplification kit is good in primer specificity, can tolerate a wide range of temperatures and is highly adaptive to subject materials.

The invention provides a fluorescence-labeled composite amplification kit for Y chromosome STR (short tandem repeat) gene loci capable of improving distinguishing capability. When the kit is used for detecting DNA (deoxyribonucleic acid) gene, not only can 17 STR gene loci of DYS391, DYS389I/II, DYS439, DYS438, DYS456, DYS458, DYS437, DYS635, DYS448, Y-GATA-H4, DYS19, DYS392, DYS393, DYS390 and DYS385a/b, which can be analyzed by the commercial kit, be amplified and analyzed, but also at least one of STR gene loci of DYS449, DYS527a/b, DYS522, DYS388, DYS447 and DYS444 can be simultaneously amplified and analyzed, so that the accumulative individual distinguishing capability and cumulative probability of exclusion of the system are improved, and the individual distinguishing capability is improved overall.

The invention relates to a composite amplification kit of 47 human autosome and Y chromosome loci and an application thereof. The invention provides a composite amplification system of the 47 human autosome and Y chromosome loci, which comprises specific primers for amplifying the 47 loci, wherein the 47 loci comprise 19 autosome STR loci, 27 Y chromosome STR loci and 1 sex recognition locus. The47 pairs of specific primers are subjected to grouping fluorescence labeling by utilizing a six-color fluorescence labeling technology, and the simultaneous high-efficiency, specific and sensitive amplification of the 47 human autosome and Y chromosome loci is achieved through the design and optimization of primer sequences and working concentrations. The detection result of the composite amplification system has high individual identification capability and good data compatibility, and can be used for paternity identification and individual identification in practice, so that the detection cost of human DNA typing is effectively reduced, and the detection working efficiency is improved.

The invention discloses a multiplex amplification detection kit for detecting 60 loca of autosomes and Y-chromosomes simultaneously. The 60 loca are amplified in a multiplex manner through a polymerase chain reaction, and amplification products of the loca are detected by using a gene sequencer. The kit can be used for detecting the 60 loca of the autosomes and the Y-chromosomes simultaneously, which is a case that most STR loca can be detected by a primary reaction with a capillary electrophoresis method at present, and databases can be built for autosome STR and Y-chromosome STR simultaneously by the primary reaction. The kit has strong adaption to biomaterials, namely, one kit can be used for amplifying various biomaterial samples, wherein different biomaterial samples comprise a male genome DNA extracted by a Chelex100 method, a magnetic bead extracting method or an organic extracting method and male blood or oral cells of human, which is/are collected by any one carrier of filterpaper, an FTA card, a cotton bud, gauze and the like. The kit has higher amplification specificity and higher thermal tolerance.

The invention discloses a Y-chromosome STR gene locus fluorescence labelled multiplex amplification kit with reinforcing resolution capability. The kit comprises the following specific oligonucleotides amplification primer pairs of 27 Y-chromosome STR gene loci: DYS456, DYS458, DYS437, DYS439, DYS392, DYS385a/b, DYS393, DYS391, DYS390, DYS635, DYS438, DYS389I, DYS19, GATA-H4, DYS389II and DYS448, wherein the kit is characterized by additionally comprising at least one of the specific oligonucleotides amplification primer pairs of 10 Y-chromosome STR gene loci: DYS576, DYS570, DYS481, DYS627, DYS460, DYS449, DYS533, DYS518 and DYF387S1. The number of the detection loci of the kit is greater than that of the market similar product both at home and abroad, so that the discrimination power and the combined power of exclusion of the system are greatly improved and the individual discriminability is generally improved. Simultaneously the invention creatively comprises 7 rapid mutation sites, so that the discrimination capacity of the close relative males can be increased, the practicability and the functionality of the kit are greatly increased, and a vicious cycle of blindly increasing the similar detection sites by the domestic kit developers is avoided.

The invention discloses a human STRtyper PCR enlarged fluoroscopic examination reagent kit with quick speed detecting, accuracy, good repeatability, good versatility, low price and practicality. The applicability of the reagent kit for Chinese is remarkable in particular. The serial products of the invention comprise the three following kits: (1) a STRtyper-20G kit with the largest STR gene locus of the global and most powerful recognizing ability; (2) a STRtyper-16GC kit which is more suitably used to build a criminal DNA database of Chinese population and identify the identity; (3) a STRtyper-10F / G kit with brand new STR gene locus. Each kit product comprises the component of 1.1 ml * 1 tube of STR PCR reaction mixture; 0.55 ml * 1 tube of STRtyper primer mixture; 120 microlitre * 1 tube of HS-Taq DNA polymerase; 0.3ml * 1 tube of STR control DNA; 50 microlitre * 1 tube of STRtyper allelomerphic gene mixture; 0.5 ml * 1 tube of internal standard; 30 microlitre* 1 tube of 310 spectrum correcting standard; 30 microlitre * 1 tube of 3100 spectrum correcting standard.

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.

The invention discloses a fluorescence labeling composite amplification kit for human Y chromosome 27 STR gene loci, which can simultaneously amplify 27 Y chromosome STR gene loci (including 24 low-mutation-rate Y chromosome STR gene loci and 3 high-mutation-rate Y chromosome STR gene loci) in a single multiplex reaction. By designing the unique gene locus configuration mode and specific primer sequences, the 27 gene loci are divided into 4 groups which are used for different fluorescence labelings. The composite amplification system has the advantages of wide temperature resistance range, high adaptability of the detection material, and high identification capacity for irrelevant and close relative male individuals.

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 discloses a DNA library construction method for high-throughput sequencing. The DNA library construction method includes the steps of carrying out multiple PCR (polymerase chain reaction) to a DNA template obtained from a to-be-tested sample through multiple pairs of fusion primers, then recycling PCR products to obtain a sequencing library. The multiple pairs of fusion primers respectively aim at different target fragments of the DNA template, and each fusion primer comprises specificity primer sequence aiming at the target fragment and linker sequence for sequencing. The multiple PCR is carried out on reaction conditions: reacting at 95 DEG C for 2 minutes; reacting within 38 cycles, wherein each cycle includes processes of preserving at the temperature of 95 DEG C for 30 seconds, cooling to the temperature from 76 DEG C to 55-58 DEG C slowly at the speed of 0.1 DEG C each second, holding the temperature for 20 seconds once cooling to the temperature of 58 DEG C, and then preserving at 72 DEG C for 30 seconds; after that, preserving at 72 DEG C for 2 minutes, and finally preserving at 4 DEG C. The invention further discloses application of the library construction method in testing STR locus and paternity identification on the basis of high-throughput sequencing.

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 Y chromosome 37 STR loci fluorescence labelled multiplex amplification kit and an application thereof. The kit comprises a specific primer for amplifying 37 Y-STR lociwhich comprise 30 low-mutation Y-STR loci and 7 rapid-mutation Y-STR loci. The kit comprises 30 low-mutation and 7 rapid-mutation rate high-polymorphism Y-STR loci, considers distinguishing of checking of a male family and different male individuals of a same male line, and is the Y-STR detection product with a maximum loci quantity on market. The primer in the kit has the advantages of strong specification, high sensitivity, and accurate parting result, and practical case examination, DNA database construction and paternity identification requirements can be completely satisfied. The kit hasstrong check-material adaptability.

The invention provides a fluorescent multiplex amplification system of 37 Y-STR gene locuses and a Y-Indel, a kit and application thereof, belonging to the technical field of biological detection. Thekit can amplify and analyze 37 Y-STR gene locuses and a Y-Indel at the same time. Six colors of fluorescence are used for marking, the size of the amplified product is 70-490bp, the operation is simple, and very high cumulative individual discernment capability and cumulative probability of paternity exclusion are realized.

The invention discloses a Y-STR compound amplification detection kit marked by a novel fluorescent labeling method and a use method thereof. The Y-STR compound amplification detection kit performs compound amplification of 27 Y-STR loca with a compound amplification primer by adopting a polymerase chain reaction; the amplification product is detected by means of a gene sequencing instrument of a single channel or multichannel capillary tube; 27 Y-STR loca comprise 20 conventional low mutation rate loca and 7 fast mutant loca. The novel fluorescent labeling method comprises the steps of: marking a first alkaline group at the 5' end of the primer by a conventional dye; and marking the sixth alkaline group with a short excitation wavelength fluorescent dye to realize fluorescent energy transfer. The signal strength generated by the method is 3-11 times higher than that of the conventional method, and an amplification result which is more stable and balanced and higher in sensitivity can be obtained.

The invention provides a method and a system for realizing Y-STR typing on the male individual by adopting 26 Y-STR genetic loci. The method comprises the following steps: acquiring DNA of the male individual, and acquiring the genotypes of the 26 Y-STR genetic loci of the DNA, wherein the 26 Y-STR genetic loci are as follows: DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS448, DYS456, DYS458, DYS635, GATA H4, DYS438, DYS439, DYS460, DYS447, DYS527a/b, DYS617, DYS522, DYS444, DYS508 and DYS533; and acquiring the Y-STR typing result of the male individual according to the genotypes of the 26 genetic loci of the male individual. With the typing result, the operations including family checking, paternity identification, detection for the male element in male and female mixed samples, and the detection for the male element in a plurality of male mixed samples can be realized on the male individual.

The invention relates to a composite amplification kit for 25 human chromosome loci and application thereof. The invention provides a composite amplification system of the 25 human chromosome loci. The system includes specific primers for amplifying the 25 loci including 23 autosomal STR loci, 1 gender recognition locus and a Y chromosome Indel locus. The 25 pairs of specific primers are subjectedto group fluorescence labeling by the six-color fluorescent labeling technology. Through design and optimization of the primer sequence and the working primer concentration, simultaneous efficient specific sensitive amplification of the 25 human chromosome loci is achieved. A detection result of the composite amplification system has high individual recognition capability and good data compatibility, and can be used in practice for paternity testing and individual recognition, thereby effectively reducing the detection cost of human DNA typing and improving the detection working efficiency.

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 discloses a fluorescence labeling composite amplification kit for 29 STR (short tandem repeat) gene loci of human Y chromosome. The kit is characterized by comprising specific amplification primers of 29 pairs of STR gene loci, wherein the STR gene loci are DYS389I/II, DYS458, DYS19, DYS385a/b, DYS393, DYS391, DYS439, DYS635, DYS392, Y GATA H4, DYS437, DYS438, DYS448, DYS456, DYS390, DYS449, DYS576, DYS481, DYS549, DYS533, DYS570, DYS643, DYS460, DYS627, DYS387a/b and DYS518. The kit has the advantages of high accumulated individual identification rate, capability of effectively reducing risk of allele crossing the border and the like.

The invention relates to a method and a reagent for improving the rate of extraction of the DNAs of castoff cells in a case trace sample. Due to the problems of trace amount of case sample, keratinization of cells, multiple polymerase chain reaction (PCR) complex amplification of 16 short tandem repeat (STR) loci and the like, the extraction of the DNAs of castoff cells has long been a difficult point in forensic medicine. According to researches, the combination of pronase digestion, bath at 70 to 100 DEG C and polyethylene glycol(PEG)-ethanol magnetic bead combined system can help to effectively extract trace DNAs and to obtain DNA fragments with a proper length, so that the need of complex amplification of the 16 STR loci in case detection can be satisfied and the success rate of the case detection is improved. The pronase digestion and warm bath allow the castoff cells to release DNAs effectively and ensures the relatively complete structure of the DNAs and the easy combination of the DNAs with the magnetic beads in the PEG-ethanol system. The use of the PEG-ethanol combined system promotes the high-efficiency DNA adsorption of the magnetic beads, ensures the relatively complete structure of the DNA and ensures that DNA fragments with proper length can be obtained to satisfy the need for the complex amplification of the 16 STR loci in case detection.

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 belongs to the technical field of legal physical evidence, in particular to a method for identifying the source of a tumor tissue based on an Identifiler system, comprising the following steps: respectively genotyping and detecting 15 STR loci of the tumor tissue and a normal tissue Identifiler system of an object to be identified; computing the number of the shared loci and the number of the shared allelic genes of the tumor tissue and the normal tissue of the object to be identified; substituting the number of the shared loci and the number of the shared allelic genes in a discrimination function and computing corresponding function values; judging the discrimination function computation result; and identifying based on a source identification rule of the tumor tissue. The invention overcomes the bottleneck that the source of the tumor tissue can not be identified when genetype variation of the STR loci of the tumor tissue occurs at present, and can be used for source identification for the tumor tissue when genetype variation occurs in the STR loci of the tumor tissue.