Human x-chromosome dna 19 locus multiplex amplification kit and its application

An X chromosome and multiple amplification technology, which is applied in the field of multiple amplification kits for 19 loci of human X chromosome DNA, can solve problems such as complex calculations and inability to guarantee non-linkage, achieve high sensitivity, save time for testing personnel, and reduce the number of reagents. The effect of time on the ratio of PCR components

Active Publication Date: 2016-02-24
AGCU SCIENTECH +1
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Human x-chromosome dna 19 locus multiplex amplification kit and its application
  • Human x-chromosome dna 19 locus multiplex amplification kit and its application
  • Human x-chromosome dna 19 locus multiplex amplification kit and its application

Examples

Experimental program
Comparison scheme
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 polyA 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 Identification of Sisterhood Using the Fluorescence-labeled Multiplexed Amplification Test System of 19 Loci

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

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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 present invention relates to a fluorescent marker compound amplification test system for 19 loci, in particular to a human X chromosome DNA 19 locus compound amplification kit and its application. The system can be applied to prenatal diagnosis, parental deletion and sister relationship Research on identification, father-daughter relationship identification, grandmother-granddaughter relationship identification, inter-generational identification, and population migration. Background technique [0002] The short tandem repeat locus (STR) is a commonly used genetic marker at present, which is a 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 produce hundreds of millions of genotype combinations, and the frequency of each combination in the population is very low. STR typing is the most important method for forensic individual identification and paternity testing a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap