Typing method for multicolor fluorescence composite detection of 20 X-SNP sites

A technology of composite detection and typing method, applied in the direction of biochemical equipment and methods, microbial measurement/testing, etc., to achieve the effect of improving the success rate

Active Publication Date: 2013-07-24
HEBEI MEDICAL UNIVERSITY
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Problems solved by technology

[0004] The invention provides a typing method for multi-color fluorescent composite detection of 20 X-SNP sites, which shortens the fragment range of the target DNA to be tested to 61-99bp, improves the success rate of highly degraded DNA typing, and can solve the problem of pro- Difficulties in the identification of complex kinship relationships such as ancestral relationship, half-sibling relationship, and sibling relationship

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  • Typing method for multicolor fluorescence composite detection of 20 X-SNP sites
  • Typing method for multicolor fluorescence composite detection of 20 X-SNP sites
  • Typing method for multicolor fluorescence composite detection of 20 X-SNP sites

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Experimental program
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Embodiment 1

[0028] (1) Extract the DNA of a woman to obtain a DNA solution;

[0029] (2) Amplification: Add the DNA solution obtained in step (1) to the amplification system, and then compound and amplify the DNA fragments of 20 X-SNP sites simultaneously in the amplification instrument.

[0030] The amplification system includes: PCR buffer solution, MgCl 2 , dNTPs, PCR primers, Taq DNA polymerase, template DNA; the concentrations or contents of the amplification system components are shown in Table 3:

[0031] Table 3 Concentration or content of amplification system components

[0032] Reagent volume 10×PCR buffer solution 2.5μL Mg 2+ (25mM) 5μL dNTPs (10mM) 1.5μL PCR primers (see Table 3 for concentration) 2.8μL Taq DNA polymerase (1U / μL) 1.5μL Template DNA (1-10ng / μL) 1μL Deionized water 10.7μL

[0033] The concentrations of PCR primers and extension primers are shown in Table 4.

[0034] Table 4 Concentrations of PC...

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Abstract

The invention discloses a typing method for multicolor fluorescence composite detection of 20 X-SNP sites, belonging to the technical field of medical genetic relationship identification. The typing method comprises the following steps of: (1) preparing a DNA solution; (2) amplifying; (3) purifying an amplification product; (4) extension reacting; (5) purifying an extension product; and (6) detecting and determining the extension product. By virtue of the invention, the segment range of target DNA to be detected is shortened to 61-99 bp, the success rate of highly degraded DNA typing is improved, and the difficulty of identifying of complicated genetic relationships, for example, ancestral relationship, half-sib relationship and sibship, can be overcome. The typing method has large advantages and potentials on complicated genetic relationship identification and forensic typing of highly degraded material DNA.

Description

technical field [0001] The invention relates to the technical field of medical genetic relationship identification. Background technique [0002] The X chromosome has unique genetic characteristics of sex-linked inheritance and cross-inheritance. It is very useful for solving complex genetic relationship identification such as father-daughter identification, mother-child identification, sister identification, half-sister identification, and intergenerational identification in forensic practice. Significance. At present, forensic DNA testing mainly focuses on short tandem repeats (short tandem repeats, STR) and single nucleotide polymorphisms (single nucleotide polymorphisms, SNP). In the complex amplification system, the amplified fragments of STR are generally large, which is not conducive to the analysis of degraded samples. At the same time, the mutation rate of some commonly used STR loci is high, which makes paternity testing face the risk of misjudgment. SNP has the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 丛斌李淑瑾王茜
Owner HEBEI MEDICAL UNIVERSITY
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