CN103757095B - Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain - Google Patents
Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain Download PDFInfo
- Publication number
- CN103757095B CN103757095B CN201310578684.9A CN201310578684A CN103757095B CN 103757095 B CN103757095 B CN 103757095B CN 201310578684 A CN201310578684 A CN 201310578684A CN 103757095 B CN103757095 B CN 103757095B
- Authority
- CN
- China
- Prior art keywords
- dna
- monosperm
- amplification
- individual
- mitochondrial dna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6858—Allele-specific amplification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain, the DNA content mainly solving prior art existence can not meet the requirement of routine inspection mixing seminal stain, it is impossible to provides the technical problem of individual entire genetic information.The method is realized by monosperm capture, monosperm DNA extraction, the nested amplification in mtDNA HV I district, the DNA sequence analysis of secondary amplified production and the step such as sperm enrichment and euchromosome STR detection.The present invention with there is individual characteristics monosperm mitochondrial DNA as Testing index, the seminal fluid mixture deriving from Different Individual is distinguished sperm according to individuality, the detection of row core DNA again, can successfully solve at present cannot be to an individual identification difficult problem for Different Individual source component in biased sample.Utilizing the feature that the content of monosperm Mitochondria DNA is more, can meet the requirement of round pcr detection mitochondrial DNA, the multiple sperms collecting mitochondrial DNA homotype can realize the detection to core DNA, to reach individual's identifying purpose.
Description
Technical field
A kind of method that the present invention relates to mixed stain is identified in sexual crime inspection, is exactly a kind of monosperm
Method individual in captured line mitochondrial DNA typing difference mixing seminal stain, belongs to prudence individual's test for identification applied technical field.
Background technology
In sexual crime case, victim may be simultaneously by the sex assault of two or more male, therefore to mixed
The test for identification closing speckle shows important especially.Mixed stain refers to tested sample from two or more individualities.In this type of case
The material evidence sample obtained in part is mixing seminal stain.The method that routine is taked at present is that application differential lysis removes women composition
After carry out euchromosome STR typing, by the calculating of peak height and the peak area at mixing spectral pattern allelic peak and combine each
The frequency of locus allele speculates individual genome type, it is impossible to determine the whole allele belonging to each individuality；Y contaminates
The analysis result of colour solid STR can improve the individual identification ability to seminal fluid mixing sample, but loses owing to it has fathership sex-linkage
The feature passed, all male of same man system have identical Y chromosome STR type, and its genotyping result can be used for getting rid of, the most not
The purpose of the same each individuality can be realized；Unicellular capture technique can obtain unicellular sample.But single spermatid
Core DNA belong to monoploid, its DNA content can not meet the requirement of routine inspection, can not provide individual entire genetic letter
Breath.
In sum, the detection method of the most all seminal fluid biased samples being applied to more than two people all can not be accomplished and determine
The purpose of each individuality.
Summary of the invention
The present invention is for the purpose of solving the problems referred to above, and the DNA content mainly solving prior art existence can not meet routine
The requirement of inspection mixing seminal stain, it is impossible to the technical problem of individual entire genetic information is provided, and it is mixed to provide one to be precisely separating
Close the sperm of Different Individual in sample and determine the method that sperm is individual again.
For achieving the above object, the present invention uses following technical proposals: the difference of monosperm captured line mitochondrial DNA typing mixes
Method individual in seminal stain, the method is realized by lower step:
One, monosperm capture
To mix seminal stain sample smear, application LMD7000 type comes card laser microdissection system (Leica, Germany) respectively
Taking single sperm, each sperm is collected in test tube respectively.
Two, monosperm DNA extraction
1, each test tube containing monosperm adds 15 μ L 5% Chelex-100,0.2 μ L10mg/ml E.C. 3.4.21.64 (eventually
Concentration 100 μ g/ml), 0.7 μ L 1M DTT(final concentration 40mM).
2, vibration 10s.
3,56 DEG C of digestion 1h～2h.
4, vibration 10s.
5, boiling water bath hatches 8min.
6, vibration 10s.
7,13000rpm is centrifuged 2min.
8, supernatant is standby in putting 4 DEG C of refrigerators.
Three, the nested amplification in mtDNA HV I district
1, amplification for the first time
Primer sequence is:
F 5′-TAC ACA ATC AAA GAC GCC CTC-3′；
R 5′-GGA TGA GGC AGG AAT CAA AGA C-3′。
Amplification system is 5 μ L, contains: 10 × buffer 0.5 μ L, 2.5mM dNTP Mix 0.5 μ L, 10uMol
The each 0.15 μ L of Primer, Template 0.5 μ L, 1u KOD-Plus-Neo 0.3 μ L, ddH2O 2.9μL。
Cycling condition is 94 DEG C of 2min；94 DEG C of 20s, 55.5 DEG C of 30s, 68 DEG C of 40s, 35cycles；68℃ 5min.
Reaction is carried out on 9700 amplification instruments, and amplification purpose fragment is 1305bp.
2, second time amplification.
Primer sequence is:
F 5′-ACC TGA ATC GGA GGA CAA C-3′；
R 5 '-CAA AGA CAG ATA CTG CGA CAT-3 ').
Amplification system is 20 μ L, contains: 10 × buffer 2 μ L, 2.5mM dNTP Mix 2 μ L, 10uMol Primer is each
0.6 μ L, for the first time amplified production 0.5 μ L, 1u KOD-Plus-Neo1.2 μ L, ddH2O 13.1μL。
Cycling condition is 94 DEG C of 2min；94 DEG C of 20s, 55.5 DEG C of 30s, 68 DEG C of 30s, 30cycles；68℃ 5min.
Reaction is carried out on 9700 amplification instruments, and amplification purpose fragment is 954bp.
Four, the DNA sequence analysis of secondary amplified production
Sequencing analysis conventional method is carried out.
Five, sperm enrichment detects with euchromosome STR
1, multitube sperm DNA extract identical for mtDNA HV I region sequence is collected in same test tube.
2, QIAamp MinElute it is incorporated inTMIn Column
3,8000rpm is centrifuged 1min.
4, adding AW1 500 μ L, 8000rpm is centrifuged 1min.
5, adding AW2 500 μ L, 8000rpm is centrifuged 1min.
6,13000rpm is centrifuged 3min.
7, add 7 μ LAE eluents, stand 5min.
8,13000rpm is centrifuged 3min.
The PowerPlex of the eluent application Promega company obtained after 9, centrifugal16HS test kit 15 normal dyes of amplification
Color str locus seat.PCR reaction is carried out on 9700 amplification instruments.PCR amplification system and cycling condition are completed by workbook.
Amplification system is: 10 μ L(Mix 2 μ L, Primer 1 μ L, DNA eluent 7 μ L).
Cycling condition is: 96 DEG C of 2min；94 DEG C of 30s, 60 DEG C of 30s, 70 DEG C of 45s, 10cycles；90 DEG C of 30s, 60
DEG C 30s, 70 DEG C of 45s, 19cycles；60℃ 30min.
Amplified production, through AB-3130 type DNA genetic analyzer electrophoretic separation and laser scanning analysis, uses 3130Data
Collection v3.0 software collects electrophoresis information, Genemapper ID v3.2 software analysis amplified fragments size, obtains
STR genotyping result.
Beneficial effects of the present invention and feature
The invention has the beneficial effects as follows, with there is individual characteristics monosperm mitochondrial DNA as Testing index, will source
Seminal fluid mixture in Different Individual distinguishes sperm, then the detection of row core DNA according to individuality, and can successfully solve at present cannot be right
An individual identification difficult problem for Different Individual source component in biased sample.Feature is, utilizes the content of monosperm Mitochondria DNA
More feature, can meet the requirement of round pcr detection mitochondrial DNA, and the multiple sperms collecting mitochondrial DNA homotype are permissible
Realize the detection to core DNA, to reach individual's identifying purpose.
Accompanying drawing explanation
Fig. 1 is mtDNA sequencing after two semen mixture autosome PCR testing results capture with monosperm
The sperm euchromosome STR genotyping result typing collection of illustrative plates that result is identical.In figure, abscissa represents " fragment length "；Vertical coordinate represents
" fluorescence signal intensity ".
It is illustrated as two male and women to sleep together the Testing and appraisal result of real case.A is two Seminal plasma
Biased sample is 15 STR analysis results of autosome after differential lysis processes, and each gene locus can detect that 1～4
Individual allele peak, it is impossible to judge the individual source at each allele peak；B Yu C is through monosperm capture, DNA extraction, line
After mitochondrial DNA sequence is analyzed, 15 STR genotyping result of the autosome of the spermatid that mtdna sequence is identical.Confirm
Mixed semen is derived from being respectively provided with the combination of the str locus type person of B Yu C, and cracking of cases result confirms that two male dislike
Doubt people.
Detailed description of the invention
Embodiment
Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain, the method is real by lower step
Existing:
One, monosperm capture
To mix seminal stain sample smear, application LMD7000 type comes card laser microdissection system (Leica, Germany) respectively
Taking single sperm, each sperm is collected in test tube respectively.
Two, monosperm DNA extraction
1, each test tube containing monosperm adds 15 μ L 5%Chelex-100,0.2 μ L10mg/ml E.C. 3.4.21.64 (eventually
Concentration 100 μ g/ml), 0.7 μ L 1M DTT(final concentration 40mM).
2, vibration 10s.
3,56 DEG C of digestion 1h～2h.
4, vibration 10s.
5, boiling water bath hatches 8min.
6, vibration 10s.
7,13000rpm is centrifuged 2min.
8, supernatant is standby in putting 4 DEG C of refrigerators.
Three, the nested amplification in mtDNA HV I district
1, amplification for the first time
Primer sequence is:
F 5′-TAC ACA ATC AAA GAC GCC CTC-3′；
R 5′-GGA TGA GGC AGG AAT CAA AGA C-3′。
Amplification system is 5 μ L, contains: 10 × buffer 0.5 μ L, 2.5mM dNTP Mix 0.5 μ L, 10uMol
The each 0.15 μ L of Primer, Template 0.5 μ L, 1u KOD-Plus-Neo 0.3 μ L, ddH2O 2.9μL。
Cycling condition is 94 DEG C of 2min；94 DEG C of 20s, 55.5 DEG C of 30s, 68 DEG C of 40s, 35cycles；68℃ 5min.
Reaction is carried out on 9700 amplification instruments, and amplification purpose fragment is 1305bp.
2, second time amplification.
Primer sequence is:
F 5′-ACC TGA ATC GGA GGA CAA C-3′；
R 5 '-CAA AGA CAG ATA CTG CGA CAT-3 ').
Amplification system is 20 μ L, contains: 10 × buffer 2 μ L, 2.5mM dNTP Mix 2 μ L, 10uMol Primer is each
0.6 μ L, for the first time amplified production 0.5 μ L, 1u KOD-Plus-Neo 1.2 μ L, ddH2O 13.1μL。
Cycling condition is 94 DEG C of 2min；94 DEG C of 20s, 55.5 DEG C of 30s, 68 DEG C of 30s, 30cycles；68℃ 5min.
Reaction is carried out on 9700 amplification instruments, and amplification purpose fragment is 954bp.
Four, the DNA sequence analysis of secondary amplified production
Sequencing analysis conventional method is carried out.
Five, sperm enrichment detects with euchromosome STR
1, multitube sperm DNA extract identical for mtDNA HV I region sequence is collected in same test tube.
2, QIAamp MinElute it is incorporated inTMIn Column
3,8000rpm is centrifuged 1min.
4, adding AW1 500 μ L, 8000rpm is centrifuged 1min.
5, adding AW2 500 μ L, 8000rpm is centrifuged 1min.
6,13000rpm is centrifuged 3min.
7, add 7 μ LAE eluents, stand 5min.
8,13000rpm is centrifuged 3min.
The PowerPlex of the eluent application Promega company obtained after 9, centrifugal16HS test kit 15 normal dyes of amplification
Color str locus seat.PCR reaction is carried out on 9700 amplification instruments.PCR amplification system and cycling condition are completed by workbook.
Amplification system is: 10 μ L(Mix 2 μ L, Primer 1 μ L, DNA eluent 7 μ L).
Cycling condition is: 96 DEG C of 2min；94 DEG C of 30s, 60 DEG C of 30s, 70 DEG C of 45s, 10cycles；90 DEG C of 30s, 60
DEG C 30s, 70 DEG C of 45s, 19cycles；60℃ 30min.
Amplified production, through AB-3130 type DNA genetic analyzer electrophoretic separation and laser scanning analysis, uses 3130 Data
Collection v3.0 software collects electrophoresis information, Genemapper ID v3.2 software analysis amplified fragments size, obtains
STR genotyping result.
Present invention can also apply to for human body nucleated cell whole in other biased samples, such as histiocyte, the thinnest
Individual's test for identification such as born of the same parents.
Claims (1)
1. method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain, the method is to be realized by lower step
:
One, monosperm capture
To mix seminal stain sample smear, application LMD7000 type comes card laser microdissection system and takes single sperm respectively, Mei Gejing
Son is collected in test tube respectively；
Two, monosperm DNA extraction
1, each test tube containing monosperm adds 15 μ L 5%Chelex-100, the 10mg/ of 0.2 μ L final concentration 100 μ g/ml
Ml E.C. 3.4.21.64, the 1M DTT of 0.7 μ L final concentration 40mM；
2, vibration 10s；
3,56 DEG C of digestion 1h～2h；
4, vibration 10s；
5, boiling water bath hatches 8min；
6, vibration 10s；
7,13000rpm is centrifuged 2min；
8, supernatant is standby in putting 4 DEG C of refrigerators；
Three, the nested amplification in mtDNA HV I district
1, amplification for the first time
Primer sequence is:
F 5'-TAC ACA ATC AAA GAC GCC CTC-3', R 5'-GGA TGA GGC AGG AAT CAA AGA C-
3'；
Amplification system is 5 μ L, contains: 10 × buffer 0.5 μ L, 2.5mM dNTP Mix 0.5 μ L, 10uMol Primer is each
0.15 μ L, Template 0.5 μ L, 1u KOD-Plus-Neo 0.3 μ L, ddH2O 2.9μL；
Cycling condition is 94 DEG C of 2min；94 DEG C of 20s, 55.5 DEG C of 30s, 68 DEG C of 40s, 35cycles；68 DEG C of 5min, reaction is 9700
Carrying out on amplification instrument, amplification purpose fragment is 1305bp；
2, second time amplification
Primer sequence is:
F 5'-ACC TGA ATC GGA GGA CAA C-3', R 5'-CAA AGA CAG ATA CTG CGA CAT-3'；
Amplification system is 20 μ L, contains: each 0.6 μ of 10 × buffer 2 μ L, 2.5mM dNTP Mix 2 μ L, 10uMol Primer
L, for the first time amplified production 0.5 μ L, 1u KOD-Plus-Neo 1.2 μ L, ddH2O 13.1μL；
Cycling condition is 94 DEG C of 2min；94 DEG C of 20s, 55.5 DEG C of 30s, 68 DEG C of 30s, 30cycles；68 DEG C of 5min, reaction is 9700
Carrying out on amplification instrument, amplification purpose fragment is 954bp；
Four, the DNA sequence analysis of secondary amplified production, sequencing analysis conventional method is carried out；
Five, sperm enrichment detects with euchromosome STR
1, multitube sperm DNA extract identical for mtDNA HV I region sequence is collected in same test tube；
2, QIAamp MinElute it is incorporated inTMIn Column；
3,8000rpm is centrifuged 1min；
4, adding AW1 500 μ L, 8000rpm is centrifuged 1min；
5, adding AW2 500 μ L, 8000rpm is centrifuged 1min；
6,13000rpm is centrifuged 3min；
7, add 7 μ LAE eluents, stand 5min；
8,13000rpm is centrifuged 3min；
9, the eluent application Promega company obtained after centrifugalTest kit 15 normal dyeing of amplification
Str locus seat, PCR reaction is carried out on 9700 amplification instruments, and PCR amplification system and cycling condition are completed by workbook；
Amplification system is: Mix 2 μ L, Primer 1 μ L, DNA eluent 7 μ L, totally 10 μ L；
Cycling condition is: 96 DEG C of 2min；94 DEG C of 30s, 60 DEG C of 30s, 70 DEG C of 45s, 10cycles；90 DEG C of 30s, 60 DEG C of 30s, 70 DEG C
45s, 19cycles；60℃30min；
Amplified production, through AB-3130 type DNA genetic analyzer electrophoretic separation and laser scanning analysis, uses 3130Data Col
Lect ion v3.0 software collects electrophoresis information, Genemapper ID v3.2 software analysis amplified fragments size, obtains STR
Genotyping result.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310578684.9A CN103757095B (en) | 2013-10-19 | 2013-11-18 | Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310492798.1 | 2013-10-19 | ||
| CN201310492798 | 2013-10-19 | ||
| CN2013104927981 | 2013-10-19 | ||
| CN201310578684.9A CN103757095B (en) | 2013-10-19 | 2013-11-18 | Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103757095A CN103757095A (en) | 2014-04-30 |
| CN103757095B true CN103757095B (en) | 2016-08-17 |
Family
ID=50524411
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310578684.9A Expired - Fee Related CN103757095B (en) | 2013-10-19 | 2013-11-18 | Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103757095B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105713895A (en) * | 2014-12-03 | 2016-06-29 | 武汉瑞博祥生物科技有限公司 | Use of agarose in DNA extraction, DNA extraction kit and DNA extraction method |
| CN106520945A (en) * | 2016-11-07 | 2017-03-22 | 江苏苏博生物医学股份有限公司 | Next generation sequencing platform-based noninvasive target mitochondrion sequencing method |
| CN109971867A (en) * | 2019-04-25 | 2019-07-05 | 中国医科大学 | A method of identifying the mixing seminal stain from two individuals |
| CN111621552B (en) * | 2019-06-13 | 2022-05-06 | 中国科学院广州生物医药与健康研究院 | Method and system for detecting mtDNA mutation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102807976A (en) * | 2011-05-30 | 2012-12-05 | 上海源奇生物医药科技有限公司 | Rapid nucleic acid extraction kit and applications thereof |
| CN102807975A (en) * | 2011-05-30 | 2012-12-05 | 熊慧 | Method for rapidly extracting nucleic acid from biological sample |
-
2013
- 2013-11-18 CN CN201310578684.9A patent/CN103757095B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102807976A (en) * | 2011-05-30 | 2012-12-05 | 上海源奇生物医药科技有限公司 | Rapid nucleic acid extraction kit and applications thereof |
| CN102807975A (en) * | 2011-05-30 | 2012-12-05 | 熊慧 | Method for rapidly extracting nucleic acid from biological sample |
Non-Patent Citations (2)
| Title |
|---|
| PALM结合低体积扩增技术在混合斑检验中的应用;任文彦等;《证据科学》;20101231;第18卷(第1期);第102-109页 * |
| ***细胞定向捕获与分离技术初步研究;赵兴春等;《刑事技术》;20121231;第14-17页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103757095A (en) | 2014-04-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103757095B (en) | Method individual in monosperm captured line mitochondrial DNA typing difference mixing seminal stain | |
| Richlen et al. | A technique for the rapid extraction of microalgal DNA from single live and preserved cells | |
| CN108220413B (en) | Fluorescent multiplex amplification kit for combined detection of human Y chromosome STR and Indel loci and application thereof | |
| Lee et al. | Rapid direct PCR for ABO blood typing | |
| CN105177146B (en) | The fluorescence labeling composite amplification kit of 27 str locus seats of human Y-chromosome and its application | |
| Jiang et al. | VDJ-Seq: deep sequencing analysis of rearranged immunoglobulin heavy chain gene to reveal clonal evolution patterns of B cell lymphoma | |
| Rucinski et al. | Comparison of two methods for isolating DNA from human skeletal remains for STR analysis | |
| Saravanan et al. | DNA barcoding detects floral origin of Indian honey samples | |
| Han et al. | A new strategy for sperm isolation and STR typing from multi-donor sperm mixtures | |
| Sun et al. | Chromatographic fingerprint and quantitative analysis of commercial Pheretima aspergillum (Guang Dilong) and its adulterants by UPLC‐DAD | |
| Paneto et al. | Heteroplasmy in hair: differences among hair and blood from the same individuals are still a matter of debate | |
| Feng et al. | Y-chromosomal haplotyping of single sperm cells isolated from semen mixtures–a successful identification of three perpetrators in a multi-suspect sexual assault case | |
| Fazi et al. | Development of two highly sensitive forensic sex determination assays based on human DYZ1 and Alu repetitive DNA elements | |
| CN105441534A (en) | Method and system for carrying out individual identification and paternity testing on Chinese populations and individuals | |
| Albastaki et al. | Design and development of novel single multiplex system incorporating 26 rapidly mutating Y‐STRs; 26 RM Yplex | |
| CN104630383B (en) | Two-humped camel polymorphism primer and its method for screening technique and identification paternity | |
| CN106755418A (en) | A kind of extron assembles sequence measurement | |
| Montesino et al. | Analysis of body fluid mixtures by mtDNA sequencing: an inter-laboratory study of the GEP-ISFG working group | |
| Quérouil et al. | Development and characterization of polymorphic microsatellite markers in neotropical fish of the genus Apistogramma (Perciformes: Labroidei: Cichlidae) | |
| CN104232751B (en) | A kind of method and system obtaining the euchromosome STR locus genotyping result of single individuality from mixing seminal stain | |
| CN104673907B (en) | A kind of system and its detection method that STR partings are examined for high flux | |
| CN109929936A (en) | It is a kind of detect human Y-chromosome rapid mutation str locus seat fluorescence labeling composite amplification kit and application | |
| Huffman et al. | Single cell genomics applications in forensic science: Current state and future directions | |
| Laurin et al. | An accelerated analytical process for the development of STR profiles for casework samples | |
| CN106755323A (en) | A kind of kit and its SNP marks for detecting cervical cancer susceptibility |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160817 Termination date: 20191118 |