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Primer combination for detecting 617 SNPs and InDel and application of primer combination in forensic identification and genetic relationship identification
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A technology of forensic identification and kinship, applied in biochemical equipment and methods, recombinant DNA technology, measurement/inspection of microorganisms, etc., can solve problems such as inability to test STR typing and difficulties in case detection
Active Publication Date: 2020-08-07
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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However, in many cases, DNA samples extracted from biological samples will be highly degraded due to human or natural factors, and effective STR typing tests cannot be performed, which makes the detection of cases difficult
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Embodiment 1
[0111] Embodiment 1, design and preparation of primer combinations
[0112] The inventors of the present invention have discovered the primer combination through a large number of preliminary experiments and effect comparison verification.
[0113] The primer set consisted of 1234 primers. All primers of the primer combination were amplified by PCR in the same system.
[0114] Among the 1234 kinds of primers, every 2 kinds of primers form a primer pair. The primer set thus consisted of 617 primer pairs.
[0115] The target sequence of each primer pair includes a SNP or InDel. Therefore, the primer combination can be used to detect 609 SNPs and 8 InDels. 609 SNPs and 8 InDels were distributed on 23 pairs of chromosomes. The information of 609 SNPs and 8 InDels are shown in Table 1.
[0118] Chromosome 3, 28 SNPs, sequence number 68-95
[0119] Chromosome 4, 34 SNPs, numbered ...
Embodiment 2
[0180] Example 2, the establishment of a method for SNP typing using a primer combination using a next-generation sequencing platform
[0181] Standard sample: 2800M Control DNA, Promega, DD7101.
[0182] Samples for sample (sample for sample 1 and sample for sample 2): two blood collection cards, blood samples were collected from two males respectively.
[0183] 1. Extract the genomic DNA of the sample to be tested, and then dilute it with ultrapure water to a DNA concentration of 1ng / μL, which is the template solution. Take a standard sample and dilute it with ultrapure water to a DNA concentration of 1ng / μL, which is the template solution. There are three template solutions in total.
[0184] 2. PCR amplification
[0185] Take the template solution, and use the primer combination designed in Example 1 to carry out PCR amplification.
[0186] The reaction system of PCR amplification is 20 μL, which contains 1 μl template solution, 1U TaqDNA polymerase and primer mixture....
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Abstract
The invention discloses a primer combination for detecting 617 SNPs and InDel and application of the primer combination in forensic identification and genetic relationship identification. The invention provides a primer group. The primer group is composed of 1234 primers, 1234 primers are sequentially shown as sequences 1 to 1234 in the sequence table. The invention also discloses application of any one of the primer groups in preparation of a forensic identification kit. The invention also protects the application of any one of the primer groups in forensic identification. The invention alsoprotects the application of any one of the primer groups in genetic relationship identification. The amplification product length of the multiplex amplification primer group is 56-225 bp, the averageamplification product length is 176 bp, the number of sites below the average amplification product length is 422, so that the multiplex amplification primer group is more suitable for forensic degradation samples. The multiple amplification primer group is good in balance, and the detection uniformity is about 83%. The method has application value for forensic identification and genetic relationship identification.
Description
technical field [0001] The invention relates to a combination of primers for detecting 617 kinds of SNPs and InDels and its application in forensic identification and genetic relationship identification. Background technique [0002] DNAtyping detection technology based on forensic science and genetics has been widely used in the field of forensic genetics. The gold standard for individual identification in the traditional forensic science field is STR typing based on capillary electrophoresis platforms. After nearly 30 years of accumulation, this technology has been able to solve the problem of DNAtyping of routine biological samples such as blood, saliva, semen, bone, and muscle. However, in many cases, DNA samples extracted from biological samples will be highly degraded due to human or natural factors, and effective STR typing tests cannot be performed, which makes the detection of cases difficult. [0003] Single nucleotide polymorphism (Single Nucleotide Polymorphi...
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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 张驰王乐康克莱季安全叶健赵光斌
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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