63 results about "Str typing" patented technology

The present invention relates to a novel STR typing strategy, which allows the simultaneous amplification and subsequent analysis of several (e.g. eleven) polymorphic systems with amplicon sizes of less than 270 bp. Thereby, after a PCR amplification the multiplex reaction is divided into two sets of STR multiplexes and analyzed separately. This multiplex system was particularly developed and tested for use in forensic investigations, where only limited amounts of DNA or only highly degraded DNA is available, for example, when the DNA is isolated from the roots of telogen hair.

The invention provides a method utilizing magnetic nanocomposite to conveniently, quickly and efficiently purify deoxyribonucleic acid (DNA) from medical samples and other samples containing trace nucleic acid. The method comprises the following steps of: (1) taking a sample to be purified, adding the guanidinium lysis solution with the final concentration of 3-6M and the dithiothreitol with the final concentration of 0.01-0.05M, uniformly mixing, and then carrying out full pyrolysis in a water bath for 5-30min; (2) forming a mixture solution containing magnetic particle-DNA composite; (3) separating the magnetic particle-DNA composite from the mixture solution; (4) cleaning; and (5) eluting to obtain the purified DNA. The method can satisfy the requirement of extracting high-quality DNA from the medical samples, and is applied in downstream polymerase chain reaction (PCR) amplification, submarine thermal reactor (STR) somatotyping and other applications; and the method does not need centrifugation, and has quick and simple operation steps. According to the method, a kit which not only is applicable to manual operation but also can be mated with an automated instrument can be produced, so the medical sample DNA can be obtained through purification.

The invention relates to the technical field of molecular biology, and especially relates to a multiplex amplification system, and a next generation sequencing typing kit and typing method for 21 micro-haplotype sites. The 21 micro-haplotype sites in the multiplex amplification system are derived from 21 autosomes, are balanced in allele frequency, are mutually independent between the sites, and have the advantages of polymorphism and low mutation rate. The kit contains 53 PCR-amplified single-end specific primers shown in SEQ ID NO. 1-SEQ ID NO.53, and can detect the 21 microhaplotype sites in a same reaction system. The multiplex amplification system, the kit and the typing method provided by the invention can solve the technical problem that a current STR typing technology cannot obtainan ideal typing result, and allele loss may occur due to excessive number of SNP sites.

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 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 reagent kit for detecting gene loci on the basis of next generation sequencing technologies and a special primer combination of the reagent kit. The primer combination comprises 60 types of DNA (deoxyribonucleic acid) molecules shown from sequences 1 to sequences 60 in sequence tables. The reagent kit for detecting the gene loci on the basis of the next generation sequencing technologies and the primer combination have the advantages that as proved by experiments, accurate typing results can be obtained when the reagent kit is used for detecting STR (short tandem repeat) typing of the 30 gene loci in standard substances 2800M; the reagent kit has important application value.

The invention relates to a next generation sequencing typing reagent kit of 43 STR sites and a method. The reagent kit comprises a PCR amplification reagent kit, wherein the PCR amplification reagentkit comprises 79 PCR amplification single-end specific primers of the 43 STR sites, and the nucleotide sequence of the PCR amplification single-end specific primers is as shown in SEQ ID NO.1-SEQ ID NO.79. The primers in the reagent kit can enable the polymorphism of each gene locus to be increased, so that probability of exclusion and system effectiveness can be improved, and the testing result is accurate and reliable; and testing results can provide new data for development of a NGS-STR typing technique, and a new detection scheme is provided for cases of target missing person survey, complex genetic relationship identification and the like through united application of a plurality of STR sites of the reagent kits in the prior art.

The invention discloses a processing method for high-throughput sequencing information of human short tandem repeat and belongs to the field of biological detection. The method comprises the steps that sequences with a preset sequencing length in STR high-throughput sequencing information of a single chip are reserved, and first sequences are formed; the first sequences are classified into different sample folders according to sample label information, the first sequences are reclassified into different STR locus folders according to STR target fragment specific primer information, and second sequences are formed; stepped reference sequences for different STR loci are established, the second sequences are compared with sequences of corresponding STR loci in the second sequences, and third sequences with the sequence similarity greater than or equal to 90% are reserved; and the threshold value of the number of sample sequencing entries is set to be 1,000, the threshold value of the number of locus sequencing entries is set to be 50, the threshold value of the number of typing sequencing entries in the loci is set to be 5, the threshold value of the number of the typing sequencing entries in the loci and the number of the locus sequencing entries is set to be 40%, and sequences with threshold values greater than or equal to the threshold values in the third sequences are screened to obtain an STR typing result.

The invention discloses a high-throughput STR typing system capable of identifying complex consanguinity and a kit thereof, and relates to the technical field of nucleic acid detection in vitro. The high-throughput STR typing system capable of identifying complex consanguinity comprises PCR primers used for amplifying 60, 118 or 179 linked autosomal STR loci; and the kit comprises a PCR primer composition, an Index connector sequence, an IGT-EM707 polymerase mixture, an amplification buffer enhancer NB, a YF buffer solution B, a database setting-up reagent and the like. The high-throughput STRtyping system capable of identifying complex consanguinity and the kit thereof provided by the invention are capable of realizing single-tube amplification of 60, 118 or 179 linked autosomal STR loci; and moreover, the STR typing system and the kit thereof are good in balance, high in sensitivity, good in specificity, and accurate in typing result. Thus, the high-throughput STR typing system capable of identifying complex consanguinity and the kit thereof can be used for identifying complex relationships of different consanguinity at the same level.

The invention discloses a system and a method for detecting an STR subtype at high throughput. The method comprises the steps of (S100) extracting a sample genome DNA; (S200) preparing an STR multiplex amplification bio-mixed system based on high throughput sequencing to carry out multiplex amplification on an STR gene locus of the sample genome DNA; and (S300) detecting the genotype of the STR gene locus by using a high throughput sequencing technology to recognize and authenticate individuals. The method is suitable for detecting the STR subtype in the field of forensic science.

The invention relates to the field of molecular biology and particularly discloses a specific primer pair for amplifying a gene D8S1179. The specific primer pair comprises D8S1179-FP:5'-TTTGTATTTCATGTGTACATTCGTAATTC-3' and D8S1179-RP:5'-ACCTATCCTGTAGATTATTTTCACTGATTG-3'. The invention further provides an STR typing method for a loca D8S1179 based on next generation sequencing and particularly provides a primer combination for being connected with a proper adaptor. The STR typing method is applied to the STR typing of the loca D8S1179. According to the primer pair capable of amplifying the gene D8S1179, after concentrations of different sample products subjected to PCR amplification, the equivalent mixing is carried out, and a genome library is constructed according to a BIOO RAPID DNA kit; the library is input into a computer by virtue of a next generation sequencer, data is subjected to bioinformatics analysis, and the typing results such as times of repetition of NDA molecules with same length, SNP of the loca and the difference among side wings of the loca of D8S1179 can be found.

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 Y chromosome parting kit and its preparation method and application. The present invention designs three pairs of public primer pairs, optimizedly select 12 X chromosome STR loci suitable for the genetic distribution of Chinese population and designs MiniSTR primers fro part of loci, and respectively adds the three pairs of public primers to the 5' end of the MiniSTR primers to obtain 24 additional primers; the use of the inventive kit can simultaneously amplify 12 X chromosome STR loci at a single-tube, avoids the competition between these primers and the formation of heterozygosity dimers, increases the amplification efficiency of each locus, wherein, the length of amplification products can be less than the 280bp, and has a high sensitivity (as low as 30pg), a higher STR parting success rate can be obtained and the amplification efficiency of medical examiner DNA degradation samples can be improved. The inventive kit has advantages of high sensitivity, high specificity, high detection efficiency, low preparation cost and the like.

The invention discloses a multiplex system, a method for detecting unbalanced mixed samples through the multiplex system and application of the method. The multiplex system comprises primers for amplifying 18 SNP-STR (single nucleotide polymorphism-short tandem repeat) sites, each site comprises a common reverse primer and two specific forward primers for increasing mutation, and the specific sequences of the primers are as shown in the SEQ ID NO.1-54 in the specification. The SNP-STR (single nucleotide polymorphism-short tandem repeat) multiplex amplification system has the advantages that the operation process is simplified, the system can be widely used on the basis of existing forensic evidence conventional inspection equipment, mixed samples of two persons at a forensic scene are detected, the same STR (short tandem repeat) typing samples are distinguished, non-invasive prenatal diagnosis and non-invasive prenatal paternity test are realized, a new technical means is provided, andclues and evidence are provided for solving a case.

The present invention discloses a multiple PCR targeted capture typing system and kit based on a high-throughput sequencing technology, and relates to the technical field of nucleic acid in-vitro detection. The STR typing system comprises PCR primers for amplification of 58, 119, or 179 linked autosomal STR gene loci. The kit comprises PCR primer combination, Index linker sequences, an IGT-EM707 polymerase mixture, an amplification buffer enhancer NB, a YF buffer B, library building reagents, etc. The provided STR typing system and kit can achieve single-tube amplification of the 58, 119, or 179 linked autosomal STR gene loci, are good in balance, high in sensitivity, good in specificity, and accurate in typing results, and can be used to identify complex consanguinity relationships of different consanguinity relationships at a same level.

The invention claims a linked autosomal short tandem repeat (STR) typing system based on a high-throughput sequencing technology and a kit, and relates to the technical field of nucleic acid in-vitrodetection. The STR typing system includes PCR primers for amplification of 61, 121 or 179 linked autosomal STR loci, and the kit includes a PCR primer combination, an Index linker sequence, an IGT-EM707 polymerase mixture, an amplification buffer enhancing agent NB, a YF buffer liquid B and a library building reagent. The STR typing system and kit provided by the invention can realize single-tubeamplification of the 61, 121 or 179 linked autosomal STR loci, have good balance, high sensitivity, good specificity, and accurate typing results, and can be used to identify complex genetic relationships of different genetic relationships at a same level.

The invention discloses a short tandem repeat (STR) parting standard substance, which is a deoxyribonucleic acid (DNA) mixture and comprises 31 DNA segments with the nucleotide sequences as 1) to 31). In the invention, all allelic genes in the bits of Chinese people groups are researched and studied through focusing on the discovered STR gene bits in the existing forensic molecular genetics, and in addition, the biostatistics technology is used for studying the gene frequency distribution of each allelic gene. On the basis, the experiments techniques such as molecular genetics, molecule biology, cell biology, genetic engineering and the like are integrally utilized, the allelic gene DNA information base is developed and built for the first time, all of the allelic gene DNA segments such as STR bits are included, further, the forensic DNA standard reference system independently developed by China is further built, and the special artificially synthesized STR standard substance is formed.

The invention discloses a DNA examination reagent for sample examination, and a special primer thereof. A DNA examination primer group provided by the invention is composed of a primer pair group used for amplifying 21 Y-STR sites, a primer pair for amplifying a sex site Amelogenin, and a primer pair for amplifying an autosome SRT site. Experiments prove that the DNA examination reagent can simultaneously examine the 21 Y-STR sites and the autosome SRT site, and the primer specificity is high, so complete SRT typing can be obtained, peak patterns are sharp and intelligible, the balance is good, no Pull-up peaks or stutter bands appear, no non-specific artificial products appear, and legal medical experts' DNA large scale sample examination requirements are completely met.

The invention provides method and system for Y-STR typing of an individual man. The method comprises the following steps: extracting the DNAs of the individual man to acquire the genotypes of 16 Y-chromosome STR gene loci of the DNAs, wherein the STR gene loci are DYS458, DYS390, DYS438, DYS392, DYS393, DYS437, DYS385a/b, GATA_H4, DYS391, DYS447, DYS19, DYS448, DYS456, DYS393, DYS635 and DYS439; and acquiring the Y-STR typing results of the individual man according to the 16 STR gene loci of the individual man. The invention also provides a system for rapid Y-STR typing of the individual man. The method and system provided by the invention can realize rapid, stable and high-quality Y-chromosome STR typing and apply the Y-STR typing results to rapid identification of patriarchal relatives, family checking and the like.

The invention discloses an analysis method for multiplex amplification STR data. The analysis method comprises the following steps: 1, site reference information: inputting data; 2, quality control (preprocessing): filtering data; 3, an STR typing method: a) searching for optimal matching, and b) referencing internal reference data. According to the method, high typing accuracy and timeliness can be guaranteed, CODIS system automatic conversion of STR loci can be achieved, the consistency of the STR loci and capillary electrophoresis data reaches up to 100% in the aspect of accuracy, and the analysis time is obviously shorter than that of mainstream analysis software in the market in the aspect of timeliness. Therefore, as an important supplementary method for whole genome sequence analysis, the process has great practical significance to forensic genetics.

The invention discloses a positive reference for STR typing. The positive reference for STR typing is prepared according to STR gene locus sites discovered in forensic molecular genetics. Experimental results show that the positive reference for STR typing can totally replace human genome DNA of 9948 for STR typing detection. Thus, the positive reference for STR typing can be used as a standard substance for a plurality of commercial kits and has important application value.