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Method for detecting noninvasive paternity tests through high-throughput sequencing

A paternity testing, high-throughput technology, applied in the field of high-throughput sequencing detection non-invasive paternity testing, can solve problems such as immeasurable development prospects, and achieve the effect of low cost, accurate results and high throughput

Inactive Publication Date: 2017-02-15
JIANGSU SUPERBIO LIFE SCI CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the application of next-generation sequencing in tumor diagnosis, non-invasive prenatal genetic screening, newborn genetic disease screening, cardiovascular disease and other genetic diseases, and forensic science is becoming more and more common, and its development prospects are immeasurable

Method used

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  • Method for detecting noninvasive paternity tests through high-throughput sequencing
  • Method for detecting noninvasive paternity tests through high-throughput sequencing
  • Method for detecting noninvasive paternity tests through high-throughput sequencing

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Embodiment 1

[0023] A method for high-throughput sequencing detection of non-invasive paternity testing, comprising the following steps:

[0024] Step a, extracting genomic DNA;

[0025] (1) Extract fetal cell-free DNA

[0026] 1) Take out 600μl plasma sample and place it at room temperature. After the sample melts, add 10μl proteinase K immediately, shake and mix, and centrifuge briefly;

[0027] 2) Immediately add 600μl GB Mix, shake and mix, and centrifuge briefly; after 10 minutes in a 56°C water bath, take out the sample tube, dry the tube wall, leave it at room temperature for 5 minutes, and centrifuge; put the adsorption column and collection tube together, and Sample information Mark the sample number on the tube cap;

[0028] 3) Add 300 μl of -20°C pre-cooled absolute ethanol, gently invert and mix 5-7 times, let stand for 5 minutes, and centrifuge;

[0029] 4) Transfer 750 μl of the sample solution to the adsorption column tube, centrifuge at 8000 rpm for 30 seconds, discard t...

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Abstract

The invention discloses a method for detecting noninvasive paternity tests through high-throughput sequencing. The method comprises steps as follows: extracting genome DNA: extracting fetus free DNA in fresh blood of a pregnant woman, pregnant DNA and suspected father DNA; performing PCR amplification by use of an amplicon kit containing multiple SNP sites, and enriching the fetus free DNA, the pregnant DNA and the suspected father DNA respectively; performing library construction on a purified multiple PCR amplification product with a library construction kit; performing quality control on a constructed library with real-time fluorescent quantitative PCR and Agilent 2100 for preparation of online sequencing; performing sequencing: performing bioinformatics analysis on an online sequencing result with software and identifying whether the genetic relationship exists. The method has the advantages of non-invasion, safety, high throughput, low cost and the like.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for high-throughput sequencing and non-invasive paternity identification. Background technique [0002] Parentage testing refers to the determination of whether there is a biological relationship between parents and children by detecting and analyzing a series of polymorphic genetic markers. Modern paternity testing methods use DNA analysis, mainly using the second-generation genetic marker—short tandem repeat (STR) typing technology, that is, through the detection, typing and statistics of 15-20 STRs on DNA samples. To achieve the purpose of calculating the probability of parentage, if there are 3 or more STR loci that are different, parent-child relationship can be excluded. However, in the application of nearly 20 years, people found that in the STR typing technology, due to the similar sequence length of different alleles, it is difficult to distinguis...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B50/06
CPCC12Q1/6869C40B50/06C12Q2531/113C12Q2537/143
Inventor 吴卫卫赵晓芳江岩朱天媛侯正强
Owner JIANGSU SUPERBIO LIFE SCI CO LTD
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