CN104911247B - A kind of DNA testing reagents and primer special for sample investigation - Google Patents
A kind of DNA testing reagents and primer special for sample investigation Download PDFInfo
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Abstract
The invention discloses a kind of DNA testing reagents and primer special for sample investigation.The invention provides DNA to examine primer sets, by being formed for expanding the primer pair group of following 21 Y STR bits point, the primer pair for expanding sex site Amelogenin and the primer pair for expanding euchromosome STR site.The experiment proves that, DNA detection reagents provided by the invention, 21 Y STR bits points and 1 euchromosome STR site can be examined simultaneously, and primer specificity is high, can obtain complete STR partings, and peak type is sharply clear, balance is good, without Pull up peaks, stutter bands, specific artefact appearance, is fully able to the requirement for meeting the extensive Sample of forensic dna nothing but.
Description
Technical field
The present invention relates to biological technical field, more particularly to a kind of DNA testing reagents for sample investigation and special draw
Thing.
Background technology
DNA inspection technologies especially (Short Tandem Repeats) STR detection techniques are from mid-term the 1980s
Since appearance, in criminal, civil case, large-scale catastrophic failure (such as：U.S.'s the September 11th attacks, Indonesia's tsunami, Wenchuan are big
Earthquake) and mass unexpected incident in played huge effect.STR (STR), also known as microsatellite DNA
(microsatellite DNA), STR detection techniques to the polymorphism of these specific STR bit points mainly by examining
Survey, after the information of multiple corresponding STR bit points is obtained, the parallel comparison of more site informations is carried out, it is achieved thereby that individual is known
Other and paternity identification.STR detections are with its distinctive technical advantage, the always focus and core of medicolegal genetics research in decades
The heart, and it is widely used in the multiple fields such as medical jurisprudence individual identification and paternity test.Forensic DNA typing standard committee of European Union
Think, although new technology continues to bring out with new genetic marker, considerably long from now in the period of in, STR will be still main
Human specific genetic marker.
Human Y-chromosome does not recombinate in genetic process with other chromosome, and sequence structure feature can be stablized
Ground is handed down from father to son and peculiar by male, has paternal inheritance feature.Most of cases of violence and all rape cases
There is male's participation in part.Therefore, effects of the Y-STR in forensic science gradually shows.In the paternity determination of no father,
Using the paternal inheritance feature of Y chromosome str locus seat (Y-STR), can be identified by paternal relative.Reflected in mixed stain
In fixed, in the case of can not particularly being separated with two-step method, Y-STR has more advantage.In case of gang raping, Y-STR is for determining
Criminal's number also has great role.Further, since Y-STR has the characteristics of paternal inheritance, not agnate, different regions groups
Body, Y-STR distribution can reveal that race, the residence even surname of suspect to its parting there is otherness.
In recent years, as the rapid development of Protocols in Molecular Biology, increasing Y-STR are developed and utilized.1997
Year, Prinz etc. establishes Y-STR composite amplification systems for the first time, Quadruplex I is named as, respectively with two kinds of fluorescence JOE
DYS19, DYS389I, DYS389II and DYS390 are marked with FAM, and has done forensic application Journal of Sex Research.American AB I
(Applied Biosystems) company is proposed Y-Filer kits, comprising DYS456, DYS389I, DYS389II,
DYS390、DYS458、DYS19、DYS385、DYS393、DYS391、DYS439、DYS635、DYS392、YGATA-H4、
17 Y-STR sites such as DYS437, DYS438, DYS448.Promega companies of the U.S. also develop comprising DYS391,
DYS389I, DYS439, DYS389II, DYS438, DYS437, DYS19, DYS392, DYS393, DYS390, DYS385a/b etc.
The PowerPlex Y kits in 12 Y-STR sites.These Y-STR test kits examine field to play more in forensic dna
Carry out bigger effect.
With the development of micro Y chromosome STR inspection technologies, its application has the trend further expanded, such as Y-DNA
Database establishment and the quick investigation of extensive sample.However, currently existing Y chromosome DNA detection reagents are in application process
In really encounter the problem of new.Because Y-STR has the characteristics of paternal inheritance, under normal circumstances, the sample of same family
The STR genotyping results obtained are identical.This there is two subject matters in extensive checkout procedure, first, big
Phenomena such as amount can not determine that there is the sample of identical Y-STR partings whether there is sample cross contamination；After two are to determine family, also
Need to carry out euchromosome STR inspection to whole samples of corresponding family again, to realize precise alignment.Inspection will be significantly greatly increased in this
The workload of personnel is tested, reduces overall inspection efficiency, increases inspection cost.
The content of the invention
It is an object of the present invention to provide DNA to examine primer sets.
DNA provided by the invention examines primer sets, including for expanding the primer pair group and use in following 21 Y-STR sites
Primer pair in amplification euchromosome STR site；
The primer pair group for being used to expand following 21 Y-STR sites is as follows：
For expanding Y-STR sites DYS460 primer pair, it is as the primer 1 shown in sequence in sequence table 1 and sequence table
Primer 2 composition shown in middle sequence 2；
For expanding Y-STR sites DYS458 primer pair, it is as the primer 3 shown in sequence in sequence table 3 and sequence table
Primer 4 shown in middle sequence 4 forms；
For expanding Y-STR sites DYS389I or Y-STR sites DYS389II primer pair, it is by sequence in sequence table 5
Primer 6 in shown primer 5 and sequence table shown in sequence 6 forms；
For expanding Y-STR sites DYS438 primer pair, it is as the primer 7 shown in sequence in sequence table 7 and sequence table
Primer 8 shown in middle sequence 8 forms；
For expanding Y-STR sites DYS390 primer pair, it is as the primer 9 shown in sequence in sequence table 9 and sequence table
Primer 10 shown in middle sequence 10 forms；
For expanding Y-STR sites DYS392 primer pair, it is as the primer 11 and sequence shown in sequence in sequence table 11
Primer 12 in table shown in sequence 12 forms；
For expanding Y-STR sites DYS393 primer pair, it is as the primer 13 and sequence shown in sequence in sequence table 13
Primer 14 in table shown in sequence 14 forms；
For expanding Y-STR sites DYS437 primer pair, it is as the primer 15 and sequence shown in sequence in sequence table 15
Primer 16 in table shown in sequence 16 forms；
For expanding Y-STR sites DYS385a or primer pair for expanding Y-STR sites DYS385b, it is by sequence table
Primer 18 in primer 17 and sequence table shown in middle sequence 17 shown in sequence 18 forms；
For expanding Y-STR sites YGATA_H4 primer pair, it is as the primer 19 and sequence shown in sequence in sequence table 19
Primer 20 in list shown in sequence 20 forms；
For expanding Y-STR sites DYS391 primer pair, it is as the primer 21 and sequence shown in sequence in sequence table 21
Primer 22 in table shown in sequence 22 forms；
For expanding Y-STR sites DYS447 primer pair, it is as the primer 23 and sequence shown in sequence in sequence table 23
Primer 24 in table shown in sequence 24 forms；
For expanding Y-STR sites DYS19 primer pair, it is as the primer 25 shown in sequence in sequence table 25 and sequence table
Primer 26 shown in middle sequence 26 forms；
For expanding Y-STR sites DYS448 primer pair, it is as the primer 27 and sequence shown in sequence in sequence table 27
Primer 28 in table shown in sequence 28 forms；
For expanding Y-STR sites DYS456 primer pair, it is as the primer 29 and sequence shown in sequence in sequence table 29
Primer 30 in table shown in sequence 30 forms；
For expanding Y-STR sites DYS635 primer pair, it is as the primer 31 and sequence shown in sequence in sequence table 31
Primer 32 in table shown in sequence 32 forms；
For expanding Y-STR sites DYS439 primer pair, it is as the primer 33 and sequence shown in sequence in sequence table 33
Primer 34 in table shown in sequence 34 forms；
For expanding Y-STR sites DYS527a or primer pair for expanding Y-STR sites DYS527b, it is by sequence table
Primer 36 in primer 35 and sequence table shown in middle sequence 35 shown in sequence 36 forms；
The primer pair for being used to expand euchromosome STR site D18S51, it is as drawing shown in sequence in sequence table 39
Primer 40 in thing 39 and sequence table shown in sequence 40 forms.
Above-mentioned primer sets also include being used for the primer pair for expanding sex site Amelogenin；
For expanding sex site Amelogenin primer pair, it is as the primer 37 and sequence shown in sequence in sequence table 37
Primer 38 in list shown in sequence 38 forms；
The primer sets by the primer pair group for expanding following 21 Y-STR sites, for expanding euchromosome STR position
The primer pair of point and the primer pair for expanding sex site Amelogenin form.
In above-mentioned primer sets, the primer 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,
19th, 20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40 mol ratio is
0.12:0.12:0.2:0.2：0.65:0.65:0.45:0.45：0.25:0.25:0.6:0.6:0.15:0.15:0.2:0.2:
0.35:0.35:0.4:0.4:0.25:0.25:0.45:0.45:0.2:0.2:0.25:0.25:0.4:0.4:0.45：0.45：
0.60:0.60:0.80:0.80:0.12:0.12:0.25:0.25。
Each equal independent packaging of bar primer in above-mentioned each primer pair.
The mol ratio when mol ratio of above-mentioned primer is uses.
In above-mentioned primer sets, a primer in each primer pair uses fluorescence labeling；
It is described be used for expand Y-STR sites DYS460 primer 15 ' ends, for expanding Y-STR sites DYS458's
5 ' ends of primer 3, primer 5 for expanding Y-STR sites DYS389I or Y-STR sites DYS389II 5 ' ends, be used for
5 ' ends of amplification DYS438 primer 7,5 ' ends of the primer 9 for expanding DYS390 and the primer for expanding DYS392
11 5 ' ends are marked with FAM；
It is described to be used to expand 5 ' ends of DYS393 primer 13, the 5 ' ends for being used to expand DYS437 primer 15
End, 5 ' ends, the institute for being used to expand Y-STR sites DYS385a or the primer 17 for expanding Y-STR sites DYS385b
5 ' the ends for stating the primer 19 for expanding YGATA_H4 are marked with HEX.
It is described be used for expand Amelogenin primer 37 5 ' ends, it is described be used for expand DYS391 primer 215 '
End, described it is used to expand 5 ' ends of DYS447 primer 23,5 ' ends of primer 25, the institute for being used to expand DYS19
5 ' the ends for stating the primer 27 for expanding DYS448 are marked with TAMRA；
It is described to be used to expand 5 ' ends of DYS456 primer 29, the 5 ' ends for being used to expand DYS635 primer 31
End, it is described be used for expand DYS439 primer 33 5 ' ends, it is described be used for expand Y-STR sites DYS527a or for expanding
5 ' ends of Y-STR sites DYS527b primer 35,5 ' ends of the primer 39 for expanding D18S51 are marked with ROX
Note.
Each bar primer of above-mentioned primer sets is independent packaging.
It is a further object to provide a kind of DNA testing reagents.
DNA detection reagents provided by the invention, by above-mentioned primer sets, dNTP, MgCl2, BSA, archaeal dna polymerase and water group
Into.
In above-mentioned DNA detection reagents, each bar primer for being used to expand in DYS460 primer pair is in the reagent
Final concentration be 0.12 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS458 is 0.20 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS389I or DYS389II be
0.65μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS438 is 0.45 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS390 is 0.25 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS392 is 0.60 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS393 is 0.15 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS437 is 0.2 μM；
For expand DYS385a or primer pair for expanding DYS385b in end of each bar primer in the reagent
Concentration is 0.35 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding YGATA_H4 is 0.4 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS391 is 0.25 μM
Final concentration of each bar primer in the reagent in primer pair for expanding DYS447 is 0.45 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS19 is 0.2 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS448 is 0.25 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS456 is 0.4 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS635 is 0.45 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS439 is 0.6 μM；
For expand DYS527a or primer pair for expanding DYS527b in end of each bar primer in the reagent
Concentration is 0.8 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding sex site Amelogenin is equal
For 0.12 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding euchromosome STR site D18S51
It is 0.25 μM.
Third object of the present invention is to provide a kind of DNA test kits.
DNA detection kits provided by the invention, including above-mentioned primer sets or mentioned reagent.
Above-mentioned primer sets or mentioned reagent are also preparing the application that is used to detect in the product of sample or sample investigation
The scope of protection of the invention.
In above-mentioned application, the sample is isolated blood or blood collection card.
The experiment proves that the present invention is normal for 21 Y-STR sites, 1 sex site Amelogenin and 1
Chromosome STR bit point, 40 primers are designed, DNA detection reagents, Neng Goutong are formed by these primers, PCR buffer solutions and DNA enzymatic
When examine 21 Y-STR sites, 1 sex site and 1 euchromosome STR site, and primer specificity high, can obtain
Whole STR partings, and peak type is sharply clear, and balance is good, no Pull-up peaks, stutter bands, nothing but the artificial production of specificity
Thing occurs, and is fully able to the requirement for meeting the extensive Sample of forensic dna.Therefore, extensive sample Y- can be used it for
In STR checkout procedures, the autosomal STR bit point of high polymorphism can play mark and distinguish same family Different Individual sample
This effect, can realize rapid investigation and the differentiation of sample.Investigation efficiency is improved, saves inspection cost, saves Check-Out Time.
Brief description of the drawings
Fig. 1 is the STR bit point layout viewing of DNA detection reagents
Fig. 2 is that the DNA testing reagents investigated for extensive sample carry out detection sample
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material used, reagent etc., unless otherwise specified, are commercially obtained in following embodiments.
The preparation of embodiment 1, the DNA testing reagents investigated for extensive sample
1st, the determination for the DNA of the extensive sample investigation sites examined
Because Y-STR has the characteristics of paternal inheritance, under normal circumstances, the STR that the sample of same family is obtained
Genotyping result is identical.This there is two subject matters in extensive checkout procedure, first, can not largely determine
Sample with identical Y-STR partings whether there is phenomena such as sample cross contamination；After two are to determine family, it is also necessary to again right
Whole samples of corresponding family carry out euchromosome STR inspection, to realize precise alignment.
Accordingly, it is determined that following 23 of the site that the DNA for extensive sample investigation is examined：21 Y-STR sites, 1
Sex site Amelogenin, 1 euchromosome STR site D18S51.
Above-mentioned 21 Y-STR sites be respectively DYS460, DYS458, DYS389I, DYS438, DYS389II, DYS390,
DYS392、DYS393、DYS437、DYS385a/b、YGATA_H4、DYS391、DYS447、DYS19、DYS448、DYS456、
DYS635、DYS439、DYS527a/b；
Above-mentioned sex site Amelogenin, it is women sample or negative sample that can directly distinguish sample to be checked；
Above-mentioned euchromosome STR site D18S51, it is capable of the Different Individual of the same family of supplementary globe to a certain extent
Sample.
The arrangement mode of its STR bit point is as shown in Figure 1.
2nd, the site special-purpose amplification primer examined for the DNA of extensive sample investigation
Composite amplification primer is designed according to above-mentioned site, sequence is as shown in table 1.
The composite amplification primer sequence of table 1
Amplimer uses FAM, HEX, TAMRA, tetra- kinds of fluorescein marks of ROX respectively.
Wherein DYS460, DYS458, DYS389I, DYS438, DYS389II, DYS390, DYS392 amplimer are used
FAM is marked.
Wherein DYS393, DYS437, DYS385a/b, YGATA_H4 amplimer are marked with HEX.
Wherein Amelogenin, DYS391, DYS447, DYS19, DYS448 amplimer are marked with TAMRA.
Wherein DYS456, DYS635, DYS439, DYS527a/b, D18S51 amplimer are marked with ROX.
Tetra- kinds of fluoresceins of FAM, HEX, TAMRA, ROX mark at 5 ' ends of positive amplimer respectively.
3rd, the preparation for the DNA testing reagents investigated for extensive sample
DNA testing reagents for extensive sample investigation are PCR amplification system, specific as follows, final concentration below
For the final concentration in amplification system：
2 × PrimerMix is to be mixed 40 primers shown in table 1 according to the concentration used shown in table 1.
Every 10 μ L PCR amplification system is as follows：(end in DNA testing reagents of each material is dense for 2 μ L5 × PCR buffer solutions
Degree is as shown in table 2), 52 × PrimerMix of μ L (final concentration of each bar primer in DNA testing reagents is as shown in table 1), 0.2 μ L
5U/ μ L FastSTAR archaeal dna polymerases (Roche companies, article No.：12032902001), remaining supplies volume with water.
Table 2 is the component of 5 × PCR buffer solutions
PCR amplification system is to be used for the DNA testing reagents of extensive sample investigation.
Therefore, DNA testing reagents primer of final concentration, final concentration of 200 μM of dNTP, final concentration of as shown in table 1
1.6mM MgCl2, final concentration of 400 μ g/mL BSA, final concentration of 1U/ μ L archaeal dna polymerases and water composition.
The application of embodiment 2, the DNA testing reagents investigated for extensive sample
1st, sample preparation
Sample is accumulated by the daily DNA database establishments of Material Evidence Identification Center, Ministry of Public Security.Sample is papery type, is blood
Liquid capture card.
Blood collection card (being known as male) is laid into a piece of disk (being no more than 1.2mm) with 0.5mm hand-held card punch,
It is placed in 0.2mL light-wall pipes.
2nd, sample expands
The 10ul DNA testing reagents prepared by embodiment 1 are added in the above-mentioned light-wall pipe equipped with blood collection card, carried out
PCR is expanded.
PCR amplification programs are：95 DEG C, 11min；94 DEG C, 30s, 59 DEG C, 2min, 72 DEG C, 1min, 28 circulations；60 DEG C,
1h, 25 DEG C, forever.
3rd, PCR primer detects
Take the above-mentioned μ L of amplified production 1 to add 10 μ L Hi-Di formamides (ABI companies), be placed in ABI3130xl genetic analyses
Capillary electrophoresis detection is carried out in instrument, concrete operation step is with reference to product description.
Data analysis is carried out using GeneMapper ID v3.2 softwares, its testing result is as shown in Fig. 2 above-mentioned sample
(male) has obtained complete STR partings, and peak type is sharply clear, and balance is good, no Pull-up peaks, stutter bands, nothing
Non-specific artefact occurs.It is fully able to the requirement for meeting the extensive Sample of forensic dna.
Therefore, can be can be used for the DNA detection reagents of the present invention in extensive sample Y-STR checkout procedures, Gao Duo
The autosomal STR bit point of state property can play a part of marking and distinguish same family Different Individual sample, can realize
The rapid investigation of sample and differentiation.Investigation efficiency is improved, saves inspection cost, saves Check-Out Time.
Claims (8)
1. a kind of DNA primer group for sample investigation, including for expanding the primer pair group and use in following 21 Y-STR sites
Primer pair in amplification euchromosome STR site；
The primer pair group for being used to expand following 21 Y-STR sites is as follows：
For expanding Y-STR sites DYS460 primer pair, it is as sequence in the primer 1 shown in sequence in sequence table 1 and sequence table
Primer 2 composition shown in row 2；
For expanding Y-STR sites DYS458 primer pair, it is as sequence in the primer 3 shown in sequence in sequence table 3 and sequence table
Primer 4 shown in row 4 forms；
For expanding Y-STR sites DYS389I or Y-STR sites DYS389II primer pair, it is as shown in sequence in sequence table 5
Primer 5 and sequence table in primer 6 shown in sequence 6 form；
For expanding Y-STR sites DYS438 primer pair, it is as sequence in the primer 7 shown in sequence in sequence table 7 and sequence table
Primer 8 shown in row 8 forms；
For expanding Y-STR sites DYS390 primer pair, it is as sequence in the primer 9 shown in sequence in sequence table 9 and sequence table
Primer 10 shown in row 10 forms；
For expanding Y-STR sites DYS392 primer pair, it is as in the primer 11 and sequence table shown in sequence in sequence table 11
Primer 12 shown in sequence 12 forms；
For expanding Y-STR sites DYS393 primer pair, it is as in the primer 13 and sequence table shown in sequence in sequence table 13
Primer 14 shown in sequence 14 forms；
For expanding Y-STR sites DYS437 primer pair, it is as in the primer 15 and sequence table shown in sequence in sequence table 15
Primer 16 shown in sequence 16 forms；
For expanding Y-STR sites DYS385a or primer pair for expanding Y-STR sites DYS385b, it is by sequence in sequence table
Primer 18 in primer 17 and sequence table shown in row 17 shown in sequence 18 forms；
For expanding Y-STR sites YGATA_H4 primer pair, it is as the primer 19 shown in sequence in sequence table 19 and sequence table
Primer 20 shown in middle sequence 20 forms；
For expanding Y-STR sites DYS391 primer pair, it is as in the primer 21 and sequence table shown in sequence in sequence table 21
Primer 22 shown in sequence 22 forms；
For expanding Y-STR sites DYS447 primer pair, it is as in the primer 23 and sequence table shown in sequence in sequence table 23
Primer 24 shown in sequence 24 forms；
For expanding Y-STR sites DYS19 primer pair, it is as sequence in the primer 25 shown in sequence in sequence table 25 and sequence table
Primer 26 shown in row 26 forms；
For expanding Y-STR sites DYS448 primer pair, it is as in the primer 27 and sequence table shown in sequence in sequence table 27
Primer 28 shown in sequence 28 forms；
For expanding Y-STR sites DYS456 primer pair, it is as in the primer 29 and sequence table shown in sequence in sequence table 29
Primer 30 shown in sequence 30 forms；
For expanding Y-STR sites DYS635 primer pair, it is as in the primer 31 and sequence table shown in sequence in sequence table 31
Primer 32 shown in sequence 32 forms；
For expanding Y-STR sites DYS439 primer pair, it is as in the primer 33 and sequence table shown in sequence in sequence table 33
Primer 34 shown in sequence 34 forms；
For expanding Y-STR sites DYS527a or primer pair for expanding Y-STR sites DYS527b, it is by sequence in sequence table
Primer 36 in primer 35 and sequence table shown in row 35 shown in sequence 36 forms；
The primer pair for being used to expand euchromosome STR site D18S51, it is as the primer 39 shown in sequence in sequence table 39
Formed with the primer 40 shown in sequence in sequence table 40；
The primer sets also include being used for the primer pair for expanding sex site Amelogenin；
For expanding sex site Amelogenin primer pair, it is as the primer 37 shown in sequence in sequence table 37 and sequence table
Primer 38 shown in middle sequence 38 forms；
The primer sets by it is described be used for expand following 21 Y-STR sites primer pair group, it is described be used for expand autosome
The primer pair of STR bit point and the primer pair composition for being used to expand sex site Amelogenin；
The primer 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,
25th, 26,27,28,29,30,31,32,33,34,35,36,37,38,39,40 mol ratio is 0.12:0.12:0.2:0.2:
0.65:0.65:0.45:0.45：0.25:0.25:0.6:0.6:0.15:0.15:0.2:0.2:0.35:0.35:0.4:0.4:
0.25:0.25:0.45:0.45:0.2:0.2:0.25:0.25:0.4:0.4:0.45：0.45：0.60:0.60:0.80:0.80:
0.12:0.12:0.25:0.25。
2. primer sets according to claim 1, it is characterised in that：A primer in each primer pair uses fluorescence
Mark；
It is described to be used to expand 5 ' ends of Y-STR sites DYS460 primer 1, the primer 3 for expanding Y-STR sites DYS458
5 ' ends, primer 5 for expanding Y-STR sites DYS389I or Y-STR sites DYS389II 5 ' ends, for expanding
5 ' ends of DYS438 primer 7,5 ' ends of primer 9 for expanding DYS390 and the primer 11 for expanding DYS392
5 ' ends are marked with FAM；
It is described to be used to expand 5 ' ends of DYS393 primer 13,5 ' ends, the institute for being used to expand DYS437 primer 15
State for expanding DYS385a or 5 ' ends of the primer 17 for expanding DYS385b, described being used to expand YGATA_H4 primer
19 5 ' ends are marked with HEX；
It is described to be used to expand 5 ' ends of Amelogenin primer 37, the 5 ' ends for being used to expand DYS391 primer 21
End, described it is used to expand 5 ' ends of DYS447 primer 23, the 5 ' ends, described for being used to expand DYS19 primer 25
5 ' ends of the primer 27 for expanding DYS448 are marked with TAMRA；
It is described to be used to expand 5 ' ends of DYS456 primer 29,5 ' ends, the institute for being used to expand DYS635 primer 31
State the primer 33 for expanding DYS439 5 ' ends, it is described be used for expand DYS527a or the primer 35 for expanding DYS527b
5 ' ends, 5 ' ends of the primer 39 for expanding D18S51 mark with ROX.
3. primer sets according to claim 1 or 2, it is characterised in that：Each bar primer of the primer sets is independent packaging.
4.DNA testing reagents, as the primer sets described in claim 1 or 2, dNTP, MgCl2, BSA, archaeal dna polymerase and water composition.
5. DNA testing reagents according to claim 4, it is characterised in that：
The final concentration of each bar primer in the reagent being used to expand in DYS460 primer pair is 0.12 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS458 is 0.20 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS389I or DYS389II is 0.65
μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS438 is 0.45 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS390 is 0.25 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS392 is 0.60 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS393 is 0.15 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS437 is 0.2 μM；
For expand DYS385a or primer pair for expanding DYS385b in final concentration of each bar primer in the reagent
It is 0.35 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding YGATA_H4 is 0.4 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS391 is 0.25 μM
Final concentration of each bar primer in the reagent in primer pair for expanding DYS447 is 0.45 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS19 is 0.2 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS448 is 0.25 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS456 is 0.4 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS635 is 0.45 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding DYS439 is 0.6 μM；
For expand DYS527a or primer pair for expanding DYS527b in final concentration of each bar primer in the reagent
It is 0.8 μM；
Final concentration of each bar primer in the reagent in primer pair for expanding sex site Amelogenin be
0.12μM；
Final concentration of each bar primer in the reagent in primer pair for expanding euchromosome STR site D18S51 be
0.25μM。
Any described primer sets or the reagent of claim 4 or 5 in 6.DNA test kits, including claim 1-3.
7. in claim 1-3 any described primer sets or the reagent of claim 4 or 5 prepare be used to detecting sample or
Application in the product of sample investigation.
8. application according to claim 7, it is characterised in that：The sample is isolated blood or blood collection card.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410087536.1A CN104911247B (en) | 2014-03-11 | 2014-03-11 | A kind of DNA testing reagents and primer special for sample investigation |
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| CN102433374A (en) * | 2010-09-29 | 2012-05-02 | 辽宁省刑事科学技术研究所 | Y-STR locus fluorescent label multiplex amplification system and application thereof |
| CN102725422A (en) * | 2009-09-11 | 2012-10-10 | 生命科技公司 | Analysis of Y-chromosome STR markers |
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| CN102725422A (en) * | 2009-09-11 | 2012-10-10 | 生命科技公司 | Analysis of Y-chromosome STR markers |
| CN102433374A (en) * | 2010-09-29 | 2012-05-02 | 辽宁省刑事科学技术研究所 | Y-STR locus fluorescent label multiplex amplification system and application thereof |
Non-Patent Citations (3)
| Title |
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| High-throughput Y-STR typing of U.S. populations with 27 regions of the Y chromosome using two multiplex PCR assays;Schoske et al.;《Forensic Sci Int》;20040128;第139卷;107-121 * |
| Y Chromosome-specific short tandem repeats in forensic casework;Mechthild Prinza et al.;《Croatian Medical Journal》;20011231;第42卷(第3期);288-291 * |
| Y染色体STR基因座复合扩增检测系统的研究进展;陈帅锋等;《中国人民公安大学学报（自然科学版）》;20041231(第42期);第49页表1，第50页右栏2.2.1-2.2.3 * |
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