Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Multiplex amplification system for rapidly mutating Y-chromosome short tandem repeats, kit and application

A compound amplification system and short tandem repeat technology, applied in recombinant DNA technology, microbial measurement/testing, DNA/RNA fragments, etc. Less problems, to achieve the effect of high polymorphism, strong directivity, high precision

Active Publication Date: 2018-03-27
SUZHOU MICROREAD GENETICS
View PDF5 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of male sperm cell production, the Y chromosome will not recombine, so in general, it will be stably inherited to its male offspring. Therefore, in the genetic analysis of males in the same family or in the same region, the mutation rate is low, The traditional Y-filer locus with few mutation sites has obvious limitations, and the individual identification rate cannot meet the requirements of forensic science

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
  • Multiplex amplification system for rapidly mutating Y-chromosome short tandem repeats, kit and application
  • Multiplex amplification system for rapidly mutating Y-chromosome short tandem repeats, kit and application
  • Multiplex amplification system for rapidly mutating Y-chromosome short tandem repeats, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Using the compound amplification system of rapid mutation Y chromosome short tandem repeat sequence to amplify the samples of various inspection materials 1. Obtaining DNA

[0067] a. Genomic DNA was extracted by Chelex-100 method or magnetic bead method.

[0068] b. Direct amplification of whole blood

[0069] c. Direct amplification of buccal swabs

[0070] Use the Chelex-100 method to extract genomic DNA (refer to "Forensic DNA Protocol". HumanaPress, 1998); or take 0.5-5 μl of anticoagulated whole blood and place it in a 500 μl centrifuge tube, shake and mix the Chelex solution to fully suspend the Chelex, each tube Add 195 μl Chelex-100 (5%) solution and 5 μl proteinase K (20mg / ml) and shake and mix well, keep warm at 56°C for two hours or overnight, take out and shake for 2 minutes, heat in boiling water for 10 minutes, centrifuge at 13000rpm for 5 minutes, and pipette carefully Transfer 150 μl of supernatant to a new centrifuge tube.

[0071] 2. Reac...

Embodiment 2

[0086] Example 2 Distinguish family samples using the compound amplification system of rapidly mutated Y chromosome short tandem repeats

[0087] 1. Collect blood samples from the family (blood samples are provided by a genetic testing center)

[0088] 2. Take 0.5-5 μl of anticoagulated whole blood into a 500 μl centrifuge tube, shake and mix the Chelex solution to make the Chelex fully suspended, add 195 μl Chelex-100 (5%) solution and 5 μl proteinase K (20 mg / ml) to each tube and shake the mixture After incubating at 56°C for two hours or overnight, take out and shake for 2 minutes, heat in boiling water for 10 minutes, then centrifuge at 13,000 rpm for 5 minutes, and carefully transfer 150 μl of the supernatant to a new centrifuge tube.

[0089] 3. Perform PCR amplification according to the amplification conditions in Example 1.

[0090] 4. Detect on the genetic analyzer according to Example 1, the amplification effect is shown in Figure 7 , Figure 8 .

[0091] 5. The...

Embodiment 3

[0095] Example 3 Verification of anti-inhibition ability

[0096] A 1ng positive control was used for amplification, and different concentrations of humic acid were added to the system. Experiments have proved that the complex system can still be fully amplified when the system contains a high concentration of inhibitors, and most of the fragments are not affected except some fragments are inhibited. Therefore, the amplification system of the present invention has high anti-suppression ability, and is suitable for common case inspection materials.

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 belongs to the technical field of biology, and relates to a multiplex amplification system for rapidly mutating Y-chromosome short tandem repeats, a kit and an application. The multiplexamplification system comprises 16 pairs of primers, and can be used for amplifying 16 STR loci: DYS630, DYS464, DYF403S1b, DYF399S1, DYS518, DYF403S1a, DYS527, DYS713, DYS612, DYS626, DYS627, DYS526,DYF404S1, DYF387S1, DYS449, and DYS547. Compared with prior art, the system is advantageous in that: 1. the system contains 16 human male Y-chromosome high-mutation rate STR, and the system is uniqueand novel with a large amount of information content and good compatibility; 2. the system has wide application range, good directivity, and high precision, and the system is suitable for legal medical expert DNA analysis in material evidence cases which relate to all cells of male; wherein biological testing materials which contain mal cells, such as bloodstain, seminal stain, saliva, hair, nail, cartilage and other human tissues, can be identified. 3. the system has good system specificity and good stability, multiple repeat checking is not needed, non-specific amplification products are not generated, and signal intensity is stable; 4. the system has high sensitivity, and DNA template amount which is low to 15pg can be accurately classified.

Description

technical field [0001] The invention belongs to the field of biotechnology and contains 16 high mutation rate Y-STR sites, 7 of which belong to multi-copy Y-STR sites, and the whole system can amplify 26 fragments. This system uses fluorescently labeled primers to simultaneously amplify multiple human male genome Y chromosome DNA short tandem repeats (STRs), efficiently and accurately perform STR typing, and then sensitively and efficiently identify forensic individuals and kinship identification, population genetic analysis, disease Gene mapping studies provide a basis for judgment. Background technique [0002] Human genome STR (short tandem repeat sequence) is a DNA genetic marker formed by tandem repeats with a few bases as the core unit. The distinction between different races, different populations and even different individuals is through the difference in core unit sequence and repeat number, which also constitutes the genetic polymorphism of STR. In the genome, th...

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
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 于在亮杨凡陈初光庞蒙维郑玉
Owner SUZHOU MICROREAD GENETICS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com