CN104911247A - DNA examination reagent for sample examination, and special primer thereof - Google Patents
DNA examination reagent for sample examination, and special primer thereof Download PDFInfo
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Abstract
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.
Description
Technical field
The present invention relates to biological technical field, particularly relate to a kind of DNA testing reagent for sample investigation and primer special.
Background technology
DNA inspection technology especially (Short Tandem Repeats) STR detection technique, since 20th century, the mid-80 came out, has all played huge effect in criminal, civil case, large-scale catastrophic failure (as: U.S.'s the September 11th attacks, Indonesia's tsunami, Wenchuan violent earthquake) and mass unexpected incident.STR (STR), also known as microsatellite DNA (microsatellite DNA), STR detection technique is mainly by detecting the polymorphism in these specificity STR sites, after the information obtaining multiple corresponding STR site, carry out the parallel comparison of multidigit dot information, thus achieve individual recognition and paternity identification.STR detects with its distinctive technical superiority, is focus and the core of medicolegal genetics research in decades always, and is widely used in multiple fields such as medical jurisprudence individual recognition and paternity test.Forensic DNA typing standard committee of European Union is thought, although new technology and new genetic marker continue to bring out, within period quite long from now on, STR will remain topmost human specific genetic marker.
Human Y-chromosome is not recombinated with other karyomit(e) in genetic process, and sequence structure feature can stably be handed down from father to son and by the male sex peculiar, there is paternal inheritance feature.Male participation is had in most of case of violence and all rape offenders.Therefore, the effect of Y-STR in forensic science manifests gradually.When there is no the paternity determination of father, utilizing the paternal inheritance feature of Y chromosome str locus seat (Y-STR), can be identified by paternal relative.In mixed stain qualification, when particularly cannot be separated by two-step approach, Y-STR has more advantage.In gang-rape case, Y-STR is for determining that criminal's number also has great role.In addition, because Y-STR has patroclinous feature, the colony of not agnate, different areas, the distribution of Y-STR also exists otherness, can disclose the race of suspect, residence or even surname to its somatotype.
In recent years, along with the develop rapidly of Protocols in Molecular Biology, increasing Y-STR is developed and utilizes.1997, the first time such as Prinz established Y-STR composite amplification system, called after Quadruplex I, marks DYS19, DYS389I, DYS389II and DYS390 respectively, and done forensic application Journal of Sex Research with two kinds of fluorescence JOE and FAM.American AB I (Applied Biosystems) company is proposed Y-Filer test kit, comprises 17 Y-STR sites such as DYS456, DYS389I, DYS389II, DYS390, DYS458, DYS19, DYS385, DYS393, DYS391, DYS439, DYS635, DYS392, YGATA-H4, DYS437, DYS438, DYS448.Promega company of the U.S. also develops the PowerPlex Y test kit comprising 12 Y-STR sites such as DYS391, DYS389I, DYS439, DYS389II, DYS438, DYS437, DYS19, DYS392, DYS393, DYS390, DYS385a/b.These Y-STR test kits play increasing effect in forensic dna inspection field.
Along with the development of micro-Y chromosome STR inspection technology, its range of application has the trend expanded further, as the quick investigation of Y-DNA database establishment and extensive sample.But current existing Y chromosome DNA detection reagent really encounters new problem in application process.Because Y-STR has patroclinous feature, under normal circumstances, the STR genotyping result that the sample of same family obtains is identical.Just there are two subject matters in this, one is to determine whether the sample with identical Y-STR somatotype exists the phenomenons such as sample cross contamination in a large number in extensive checkout procedure; Two is after determining family, also needs again to carry out euchromosome STR inspection, to realize precise alignment to whole samples of corresponding family.This will greatly increase the workload of reviewer, reduces overall inspection efficiency, increases inspection cost.
Summary of the invention
An object of the present invention is to provide DNA and check primer sets.
DNA provided by the invention checks primer sets, comprises for the primer pair group in following 21 Y-STR sites of increasing and the primer pair for euchromosome STR site of increasing;
The described primer pair group for following 21 Y-STR sites of increasing is as follows:
For the primer pair of the Y-STR site DYS460 that increases, it is made up of the primer 2 shown in sequence 2 in the primer 1 shown in sequence in sequence table 1 and sequence table;
For the primer pair of the Y-STR site DYS458 that increases, it is made up of the primer 4 shown in sequence 4 in the primer 3 shown in sequence in sequence table 3 and sequence table;
For the primer pair of increase Y-STR site DYS389I or Y-STR site DYS389II, it is made up of the primer 6 shown in sequence 6 in the primer 5 shown in sequence in sequence table 5 and sequence table;
For the primer pair of the Y-STR site DYS438 that increases, it is made up of the primer 8 shown in sequence 8 in the primer 7 shown in sequence in sequence table 7 and sequence table;
For the primer pair of the Y-STR site DYS390 that increases, it is made up of the primer 10 shown in sequence 10 in the primer 9 shown in sequence in sequence table 9 and sequence table;
For the primer pair of the Y-STR site DYS392 that increases, it is made up of the primer 12 shown in sequence 12 in the primer 11 shown in sequence in sequence table 11 and sequence table;
For the primer pair of the Y-STR site DYS393 that increases, it is made up of the primer 14 shown in sequence 14 in the primer 13 shown in sequence in sequence table 13 and sequence table;
For the primer pair of the Y-STR site DYS437 that increases, it is made up of the primer 16 shown in sequence 16 in the primer 15 shown in sequence in sequence table 15 and sequence table;
For Y-STR site DYS385a or the primer pair for the Y-STR site DYS385b that increases of increasing, it is made up of the primer 18 shown in sequence 18 in the primer 17 shown in sequence in sequence table 17 and sequence table;
For the primer pair of the Y-STR site YGATA_H4 that increases, it is made up of the primer 20 shown in sequence 20 in the primer 19 shown in sequence in sequence table 19 and sequence table;
For the primer pair of the Y-STR site DYS391 that increases, it is made up of the primer 22 shown in sequence 22 in the primer 21 shown in sequence in sequence table 21 and sequence table;
For the primer pair of the Y-STR site DYS447 that increases, it is made up of the primer 24 shown in sequence 24 in the primer 23 shown in sequence in sequence table 23 and sequence table;
For the primer pair of the Y-STR site DYS19 that increases, it is made up of the primer 26 shown in sequence 26 in the primer 25 shown in sequence in sequence table 25 and sequence table;
For the primer pair of the Y-STR site DYS448 that increases, it is made up of the primer 28 shown in sequence 28 in the primer 27 shown in sequence in sequence table 27 and sequence table;
For the primer pair of the Y-STR site DYS456 that increases, it is made up of the primer 30 shown in sequence 30 in the primer 29 shown in sequence in sequence table 29 and sequence table;
For the primer pair of the Y-STR site DYS635 that increases, it is made up of the primer 32 shown in sequence 32 in the primer 31 shown in sequence in sequence table 31 and sequence table;
For the primer pair of the Y-STR site DYS439 that increases, it is made up of the primer 34 shown in sequence 34 in the primer 33 shown in sequence in sequence table 33 and sequence table;
For Y-STR site DYS527a or the primer pair for the Y-STR site DYS527b that increases of increasing, it is made up of the primer 36 shown in sequence 36 in the primer 35 shown in sequence in sequence table 35 and sequence table;
The described primer pair for the euchromosome STR site D18S51 that increases, it is made up of the primer 40 shown in sequence 40 in the primer 39 shown in sequence in sequence table 39 and sequence table.
Above-mentioned primer sets also comprises the primer pair for the sex site Amelogenin that increases;
For the primer pair of the sex site Amelogenin that increases, it is made up of the primer 38 shown in sequence 38 in the primer 37 shown in sequence in sequence table 37 and sequence table;
Described primer sets is made up of the primer pair group for following 21 Y-STR sites of increasing, the primer pair for euchromosome STR site of increasing and the primer pair for the sex site Amelogenin that increases.
In above-mentioned primer sets, described 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, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, the mol ratio of 40 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.
The equal independent packaging of each bar primer in each primer pair above-mentioned.
The mol ratio when mol ratio of above-mentioned primer is use.
In above-mentioned primer sets, a primer in primer pair described in each all uses fluorescent mark;
5 ' end of the described primer 1 for the Y-STR site DYS460 that increases, for 5 ' end of the primer 3 of the Y-STR site DYS458 that increases, 5 ' end for the primer 5 of increase Y-STR site DYS389I or Y-STR site DYS389II, the 5 ' end for the primer 7 of the DYS438 that increases, the primer 9 for the DYS390 that increases 5 ' end and all mark with FAM for 5 ' end of the primer 11 of the DYS392 that increases;
5 ' end of the described primer 13 for the DYS393 that increases, 5 ' end of the described primer 15 for the DYS437 that increases, described for increasing Y-STR site DYS385a or all mark with HEX for 5 ' end of the primer 17 of the Y-STR site DYS385b that increases, 5 ' end of the described primer 19 for the YGATA_H4 that increases.
5 ' end of the described primer 37 for the Amelogenin that increases, 5 ' end of the described primer 21 for the DYS391 that increases, 5 ' end of the described primer 23 for the DYS447 that increases, 5 ' end of the described primer 25 for the DYS19 that increases, 5 ' end of the described primer 27 for the DYS448 that increases all mark with TAMRA;
5 ' end of 5 ' end of the described primer 29 for the DYS456 that increases, 5 ' end of the described primer 31 for the DYS635 that increases, the described primer 33 for the DYS439 that increases, described for increasing Y-STR site DYS527a or all mark with ROX for 5 ' end of the primer 35 of the Y-STR site DYS527b that increases, 5 ' end of the described primer 39 for the D18S51 that increases.
Each bar primer of above-mentioned primer sets is independent packaging.
Another object of the present invention is to provide a kind of DNA testing reagent.
DNA detection reagent provided by the invention, by above-mentioned primer sets, dNTP, MgCl
2, BSA, archaeal dna polymerase and water composition.
In above-mentioned DNA detection reagent, be describedly 0.12 μM for the final concentration of each bar primer in described reagent increased in the primer pair of DYS460;
For the DYS458 that increases primer pair in the final concentration of each bar primer in described reagent be 0.20 μM;
0.65 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS389I or DYS389II;
0.45 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS438;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS390;
0.60 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS392;
0.15 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS393;
0.2 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS437;
For increasing DYS385a or be 0.35 μM for the final concentration of each bar primer in described reagent increased in the primer pair of DYS385b;
0.4 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of YGATA_H4;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS391
0.45 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS447;
0.2 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS19;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS448;
0.4 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS456;
0.45 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS635;
0.6 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS439;
For increasing DYS527a or be 0.8 μM for the final concentration of each bar primer in described reagent increased in the primer pair of DYS527b;
0.12 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of sex site Amelogenin;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of euchromosome STR site D18S51.
3rd object of the present invention is to provide a kind of DNA test kit.
DNA detection test kit provided by the invention, comprises above-mentioned primer sets or mentioned reagent.
Above-mentioned primer sets or mentioned reagent are also being the scope of protection of the invention for the preparation of the application detected in the product of sample or sample investigation.
In above-mentioned application, described sample is isolated blood or blood collection card.
Experiment of the present invention proves, the present invention is directed to 21 Y-STR sites, 1 sex site Amelogenin and 1 euchromosome STR site, design 40 primers, by these primers, PCR damping fluid and DNA enzymatic composition DNA detection reagent, 21 Y-STR sites can be checked simultaneously, 1 sex site and 1 euchromosome STR site, and primer specificity is high, complete STR somatotype can be obtained, and peak type is sharply clear, balance is good, without Pull-up peak, stutter is with, specificity artifact occurs nothing but, the requirement of the extensive Sample of forensic dna can be met completely.Therefore, can use it in extensive sample Y-STR checkout procedure, the autosomal STR site of high polymorphism can be played mark and be distinguished the effect of same family Different Individual sample, can realize rapid investigation and the differentiation of sample.Improve investigation efficiency, save inspection cost, save proving time.
Accompanying drawing explanation
Fig. 1 is the STR site layout viewing of DNA detection reagent
Fig. 2 is that the DNA testing reagent investigated for extensive sample carries out detection sample
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Embodiment 1, the preparation of DNA testing reagent investigated for extensive sample
The determination in the site that the DNA 1, investigated for extensive sample checks
Because Y-STR has patroclinous feature, under normal circumstances, the STR genotyping result that the sample of same family obtains is identical.Just there are two subject matters in this, one is to determine whether the sample with identical Y-STR somatotype exists the phenomenons such as sample cross contamination in a large number in extensive checkout procedure; Two is after determining family, also needs again to carry out euchromosome STR inspection, to realize precise alignment to whole samples of corresponding family.
Therefore, following 23: 21 Y-STR sites, site of the DNA inspection of extensive sample investigation, 1 sex site Amelogenin, 1 euchromosome STR site D18S51 is determined.
Above-mentioned 21 Y-STR sites are 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, directly can distinguish sample to be checked is women's sample or negative sample;
Above-mentioned euchromosome STR site D18S51, can the Different Individual sample of the same family of supplementary globe to a certain extent.
The arrangement mode in its STR site is illustrated in fig. 1 shown below.
2, the site special-purpose amplification primer that the DNA investigated for extensive sample checks
According to above-mentioned site design composite amplification primer, sequence is as shown in table 1.
Table 1 composite amplification primer sequence
Amplimer uses FAM respectively, and HEX, TAMRA, ROX tetra-kinds is fluorescein-labelled.
Wherein the amplimer FAM of DYS460, DYS458, DYS389I, DYS438, DYS389II, DYS390, DYS392 marks.
Wherein the amplimer HEX of DYS393, DYS437, DYS385a/b, YGATA_H4 marks.
Wherein the amplimer TAMRA of Amelogenin, DYS391, DYS447, DYS19, DYS448 marks.
Wherein the amplimer ROX of DYS456, DYS635, DYS439, DYS527a/b, D18S51 marks.
FAM, HEX, TAMRA, ROX tetra-kinds of fluoresceins are marked at 5 ' end of forward amplimer respectively.
The preparation of the DNA testing reagent 3, investigated for extensive sample
The DNA testing reagent investigated for extensive sample is PCR amplification system, specific as follows, and final concentration is below the final concentration in amplification system:
2 × PrimerMix is for mixing the primer of 40 shown in table 1 according to the concentration used table 1 Suo Shi.
The PCR amplification system of every 10 μ L is as follows: 2 μ L5 × PCR damping fluids (final concentration in DNA testing reagent of each material is as shown in table 2), the final concentration of 5 μ L2 × PrimerMix(each bar primer in DNA testing reagent are as shown in table 1), FastSTAR archaeal dna polymerase (the Roche company of 0.2 μ L5U/ μ L, article No.: 12032902001), all the other supply volume with water.
Table 2 is the component of 5 × PCR damping fluid
| PCR Buffer composition | Final concentration |
| Tris-HCl | 50mM |
| dNTP | 1mM each |
| MgCl 2 | 8mM |
| BSA | 2mg/mL |
| KCl | 250mM |
PCR amplification system is the DNA testing reagent investigated for extensive sample.
Therefore, DNA testing reagent by having the primer of final concentration shown in table 1, final concentration is 200 μMs of dNTP, final concentration is the MgCl of 1.6mM
2, final concentration is the BSA of 400 μ g/mL, final concentration is 1U/ μ L archaeal dna polymerase and water composition.
Embodiment 2, the application of DNA testing reagent investigated for extensive sample
1, sample preparation
Sample is accumulated by the daily DNA database establishment of Material Evidence Identification Center, Ministry of Public Security.Sample is papery type, is blood collection card.
Blood collection card (being known as the male sex) 0.5mm hand-held punch tool is laid a slice disk (being no more than 1.2mm), is placed in 0.2mL thin-walled tube.
2, sample amplification
The DNA testing reagent prepared by embodiment 1 by 10ul adds and is above-mentionedly equipped with in the thin-walled tube of blood collection card, carries out pcr amplification.
Pcr amplification program is: 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.
3, PCR primer detects
Get above-mentioned amplified production 1 μ L and add 10 μ L Hi-Di methane amides (ABI company), be placed in ABI3130xl genetic analyzer and carry out capillary electrophoresis detection, concrete operation step is with reference to product description.
Adopt GeneMapper ID v3.2 software to carry out data analysis, its detected result is illustrated in fig. 2 shown below, and above-mentioned sample (male sex) obtains complete STR somatotype, and peak type is sharply clear, balance is good, and without Pull-up peak, stutter band, specificity artifact occurs nothing but.The requirement of the extensive Sample of forensic dna can be met completely.
Therefore, can may be used in extensive sample Y-STR checkout procedure with DNA detection reagent of the present invention, the autosomal STR site of high polymorphism can be played mark and be distinguished the effect of same family Different Individual sample, can realize rapid investigation and the differentiation of sample.Improve investigation efficiency, save inspection cost, save proving time.
Claims (10)
1.DNA checks primer sets, comprises for the primer pair group in following 21 Y-STR sites of increasing and the primer pair for euchromosome STR site of increasing;
The described primer pair group for following 21 Y-STR sites of increasing is as follows:
For the primer pair of the Y-STR site DYS460 that increases, it is made up of the primer 2 shown in sequence 2 in the primer 1 shown in sequence in sequence table 1 and sequence table;
For the primer pair of the Y-STR site DYS458 that increases, it is made up of the primer 4 shown in sequence 4 in the primer 3 shown in sequence in sequence table 3 and sequence table;
For the primer pair of increase Y-STR site DYS389I or Y-STR site DYS389II, it is made up of the primer 6 shown in sequence 6 in the primer 5 shown in sequence in sequence table 5 and sequence table;
For the primer pair of the Y-STR site DYS438 that increases, it is made up of the primer 8 shown in sequence 8 in the primer 7 shown in sequence in sequence table 7 and sequence table;
For the primer pair of the Y-STR site DYS390 that increases, it is made up of the primer 10 shown in sequence 10 in the primer 9 shown in sequence in sequence table 9 and sequence table;
For the primer pair of the Y-STR site DYS392 that increases, it is made up of the primer 12 shown in sequence 12 in the primer 11 shown in sequence in sequence table 11 and sequence table;
For the primer pair of the Y-STR site DYS393 that increases, it is made up of the primer 14 shown in sequence 14 in the primer 13 shown in sequence in sequence table 13 and sequence table;
For the primer pair of the Y-STR site DYS437 that increases, it is made up of the primer 16 shown in sequence 16 in the primer 15 shown in sequence in sequence table 15 and sequence table;
For Y-STR site DYS385a or the primer pair for the Y-STR site DYS385b that increases of increasing, it is made up of the primer 18 shown in sequence 18 in the primer 17 shown in sequence in sequence table 17 and sequence table;
For the primer pair of the Y-STR site YGATA_H4 that increases, it is made up of the primer 20 shown in sequence 20 in the primer 19 shown in sequence in sequence table 19 and sequence table;
For the primer pair of the Y-STR site DYS391 that increases, it is made up of the primer 22 shown in sequence 22 in the primer 21 shown in sequence in sequence table 21 and sequence table;
For the primer pair of the Y-STR site DYS447 that increases, it is made up of the primer 24 shown in sequence 24 in the primer 23 shown in sequence in sequence table 23 and sequence table;
For the primer pair of the Y-STR site DYS19 that increases, it is made up of the primer 26 shown in sequence 26 in the primer 25 shown in sequence in sequence table 25 and sequence table;
For the primer pair of the Y-STR site DYS448 that increases, it is made up of the primer 28 shown in sequence 28 in the primer 27 shown in sequence in sequence table 27 and sequence table;
For the primer pair of the Y-STR site DYS456 that increases, it is made up of the primer 30 shown in sequence 30 in the primer 29 shown in sequence in sequence table 29 and sequence table;
For the primer pair of the Y-STR site DYS635 that increases, it is made up of the primer 32 shown in sequence 32 in the primer 31 shown in sequence in sequence table 31 and sequence table;
For the primer pair of the Y-STR site DYS439 that increases, it is made up of the primer 34 shown in sequence 34 in the primer 33 shown in sequence in sequence table 33 and sequence table;
For Y-STR site DYS527a or the primer pair for the Y-STR site DYS527b that increases of increasing, it is made up of the primer 36 shown in sequence 36 in the primer 35 shown in sequence in sequence table 35 and sequence table;
The described primer pair for the euchromosome STR site D18S51 that increases, it is made up of the primer 40 shown in sequence 40 in the primer 39 shown in sequence in sequence table 39 and sequence table.
2. primer sets according to claim 1, is characterized in that: described primer sets also comprises the primer pair for the sex site Amelogenin that increases;
For the primer pair of the sex site Amelogenin that increases, it is made up of the primer 38 shown in sequence 38 in the primer 37 shown in sequence in sequence table 37 and sequence table;
Described primer sets is made up of the described primer pair group for following 21 Y-STR sites of increasing, the described primer pair for euchromosome STR site of increasing and the described primer pair for the sex site Amelogenin that increases.
3. primer sets according to claim 2, is characterized in that:
Described 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, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, the mol ratio of 40 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.
4. the primer sets according to Claims 2 or 3, is characterized in that: a primer in primer pair described in each all uses fluorescent mark;
5 ' end of the described primer 1 for the Y-STR site DYS460 that increases, for 5 ' end of the primer 3 of the Y-STR site DYS458 that increases, 5 ' end for the primer 5 of increase Y-STR site DYS389I or Y-STR site DYS389II, the 5 ' end for the primer 7 of the DYS438 that increases, the primer 9 for the DYS390 that increases 5 ' end and all mark with FAM for 5 ' end of the primer 11 of the DYS392 that increases;
5 ' end of the described primer 13 for the DYS393 that increases, 5 ' end of the described primer 15 for the DYS437 that increases, described for increasing DYS385a or all mark with HEX for 5 ' end of the primer 17 of the DYS385b that increases, 5 ' end of the described primer 19 for the YGATA_H4 that increases.
5 ' end of the described primer 37 for the Amelogenin that increases, 5 ' end of the described primer 21 for the DYS391 that increases, 5 ' end of the described primer 23 for the DYS447 that increases, 5 ' end of the described primer 25 for the DYS19 that increases, 5 ' end of the described primer 27 for the DYS448 that increases all mark with TAMRA;
5 ' end of 5 ' end of the described primer 29 for the DYS456 that increases, 5 ' end of the described primer 31 for the DYS635 that increases, the described primer 33 for the DYS439 that increases, described for increasing DYS527a or all mark with ROX for 5 ' end of the primer 35 of the DYS527b that increases, 5 ' end of the described primer 39 for the D18S51 that increases.
5., according to described primer sets arbitrary in claim 1-4, it is characterized in that: each bar primer of described primer sets is independent packaging.
6.DNA testing reagent, by described primer sets arbitrary in claim 1-5, dNTP, MgCl
2, BSA, archaeal dna polymerase and water composition.
7. DNA testing reagent according to claim 6, is characterized in that:
Describedly be 0.12 μM for the final concentration of each bar primer in described reagent increased in the primer pair of DYS460;
For the DYS458 that increases primer pair in the final concentration of each bar primer in described reagent be 0.20 μM;
0.65 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS389I or DYS389II;
0.45 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS438;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS390;
0.60 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS392;
0.15 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS393;
0.2 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS437;
For increasing DYS385a or be 0.35 μM for the final concentration of each bar primer in described reagent increased in the primer pair of DYS385b;
0.4 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of YGATA_H4;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS391
0.45 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS447;
0.2 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS19;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS448;
0.4 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS456;
0.45 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS635;
0.6 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of DYS439;
For increasing DYS527a or be 0.8 μM for the final concentration of each bar primer in described reagent increased in the primer pair of DYS527b;
0.12 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of sex site Amelogenin;
0.25 μM is for the final concentration of each bar primer in described reagent increased in the primer pair of euchromosome STR site D18S51.
8.DNA test kit, to comprise in claim 1-5 reagent described in arbitrary described primer sets or claim 6 or 7.
9. in claim 1-5 reagent described in arbitrary described primer sets or claim 6 or 7 for the preparation of the application detected in the product of sample or sample investigation.
10. application according to claim 9, is characterized in that: described sample is isolated blood or blood collection card.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520980A (en) * | 2016-11-30 | 2017-03-22 | 公安部物证鉴定中心 | Method and system for Y-STR typing of individual man |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
-
2014
- 2014-03-11 CN CN201410087536.1A patent/CN104911247B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
|---|
| MECHTHILD PRINZA ET AL.: "Y Chromosome-specific short tandem repeats in forensic casework", 《CROATIAN MEDICAL JOURNAL》 * |
| SCHOSKE ET AL.: "High-throughput Y-STR typing of U.S. populations with 27 regions of the Y chromosome using two multiplex PCR assays", 《FORENSIC SCI INT》 * |
| 陈帅锋等: "Y染色体STR基因座复合扩增检测系统的研究进展", 《中国人民公安大学学报（自然科学版）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520980A (en) * | 2016-11-30 | 2017-03-22 | 公安部物证鉴定中心 | Method and system for Y-STR typing of individual man |
| CN106520980B (en) * | 2016-11-30 | 2019-11-05 | 公安部物证鉴定中心 | A kind of pair of male individual carries out the method and system of Y-STR parting |
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| CN104911247B (en) | 2018-03-23 |
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