Human X-chromosomal DNA 19-gene-locus multiplex amplification reagent kit and application

An X-chromosome and compound amplification technology is applied in the field of compound amplification kits for 19 loci of human X-chromosome DNA, which can solve the problems of complex calculation and inability to guarantee non-linkage, achieve high sensitivity, save the time of testing personnel, and reduce the number of The effect of time than PCR components

Active Publication Date: 2014-09-24
AGCU SCIENTECH +1
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  • Claims
  • Application Information

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Problems solved by technology

Although there are 17 X-STR loci in Beijing's basic cognition, the loci it selects come from different linkage groups. Since the length of the X chromosome is 155Mb, the entire chromosome has four linkage groups that can guarantee that they are not linked to each other. Any additional sites cannot be guaranteed to be unlinked, and the calculation of the final result is complicated

Method used

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  • Human X-chromosomal DNA 19-gene-locus multiplex amplification reagent kit and application
  • Human X-chromosomal DNA 19-gene-locus multiplex amplification reagent kit and application
  • Human X-chromosomal DNA 19-gene-locus multiplex amplification reagent kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1 Determination of loci, primer design and establishment of kit amplification parameters

[0036] The selected loci mainly consider the following factors: 1. Polymorphism, the GD value of the selected STR loci is relatively high in the Chinese population > 0.6, 2. Select closely linked loci in different linkage groups, located in the same linkage group The loci are inherited in linkage and can be used to observe polymorphisms by haplotype. 3. Select loci with a relatively low mutation rate, such as DXS8377 and DXS10011 with a mutation rate of 3.7% and 2.5%, which cannot be selected. 4. Consider primer design factors, such as the DXS10148 template sequence, as follows, there are 14 poly A structures not far before the core repeat region, so the primer cannot span this region. Primers can only be in the back region, such as AAAAGGGGGAAGGAAGGAAG. The primer contains 3 repeats of GGAA, which is enough to match the locus of TTCC in the core repeat region to produce...

Embodiment 2

[0062] Example 2 Application of 19 loci fluorescent labeling compound amplification test system for sister relationship identification

[0063] like image 3 , linked X loci have unique advantages in complex paternity testing, such as sisterhood determination. B and A may be half-sisters, and their biological father M has died, M1 and M2 are M's siblings, and M3 is M's sister.

[0064] 1. Collect blood samples in identification cases: The samples in this experiment were provided by XX paternity testing institutions. DNA extraction Using the Chelex-100 method to extract genomic DNA from two whole blood samples: take 0.5-5 μl of whole blood into a sterilized 1.5 ml centrifuge tube, add sdH 2 Put O1ml in the tube, shake for a few seconds; after 10 minutes at room temperature, shake for a few seconds, centrifuge at 12,000 rpm for 3 minutes, discard the supernatant, keep enough supernatant to cover the precipitate, do not stir the precipitate; add 200 μl of 5% Chelex-100 , shake...

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Abstract

The invention discloses a human X-chromosomal DNA 19-gene-locus multiplex amplification reagent kit and application. The reagent kit can detect 19 X-chromosomal gene loca at the same time. According to the reagent kit, the 19 gene loca fall into four groups, and five-color fluorescence markers are involved together; by the adoption of the reagent kit, the loca are compatible with a current international common reagent kit, a linkage group with the higher research basis in the Chinese population is added, and the reagent kit is effectively applied into the aspects of antenatal diagnosis, relationship identification of sisters without parents, father-daughter relationship identification, grandmother-granddaughter relationship identification, atavistic relative identification, population migration research and the like. According to the human X-chromosomal DNA 19-gene-locus multiplex amplification reagent kit and application, a fluorescence marker multiplex amplification system is high in sensitivity, and all the 19 gene loca can be detected under the condition that the DNA template amount is 0.15 ng.

Description

technical field [0001] The invention relates to a fluorescent labeling compound amplification test system for 19 loci, in particular to a compound amplification kit for 19 loci of human X chromosome DNA and its application. The system can be applied to prenatal diagnosis, parental deletion and sister relationship Research on identification, identification of father-daughter relationship, identification of relationship between grandmother and granddaughter, identification of relatives across generations and population migration. Background technique [0002] Short tandem repeat locus (STR) is a commonly used genetic marker, which is 2-6 bp long tandem repeat sequence, and its length is between tens to hundreds of bp. The DNA sequence formed by this tandem repeat can generate hundreds of millions of genotype combinations, and the frequency of each combination in the population is very low. STR typing is the most important for forensic individual identification and paternity te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 郑卫国陈林丽李科杰卢青葛斌文吴微微郝宏蕾任文彦周怀谷吕德坚
Owner AGCU SCIENTECH
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