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Method and kit for extracting genomic DNA from whole blood

A technology of genome and kit, applied in the field of nucleic acid

Inactive Publication Date: 2019-04-12
BEIJING USCI MEDICAL LAB CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In summary, the existing whole blood genomic DNA extraction methods are difficult to solve the problem of false positives in the detection results of the increase or decrease in the number of copies of a large number of fragments of multiple chromosomes

Method used

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  • Method and kit for extracting genomic DNA from whole blood
  • Method and kit for extracting genomic DNA from whole blood
  • Method and kit for extracting genomic DNA from whole blood

Examples

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Embodiment 1

[0053] This embodiment selects three normal people's plasma samples (numbered 1, 2, and 3), and each sample is divided into two groups, one group uses the method of the application (group A), and one group uses the existing method (group B) , detailed steps will not be listed here), the genomic DNA of leukocytes was extracted from whole blood, and sequenced and bioinformatics analysis was performed after routine library construction, focusing on the chromosomal abnormalities of the two groups of normal samples. Among them, in group A, the specific process of the extraction method of this application is as follows:

[0054] 1. Extraction method of leukocyte genomic DNA

[0055] (1) Lysis of whole red blood cells

[0056] First add 0.5mL of fresh or frozen whole blood and 1.0mL of Buffer A to a 1.5mL centrifuge tube (the specific formula is: 10mmol / L Tris-HCL pH7.6, 10mmol / L NaCl, 5mmol / L MgCl 2 ), invert and mix 10 times and then stand at room temperature for 10 min. Put the...

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Abstract

The present invention provides a method and kit for extracting genomic DNA from whole blood. The method comprises: separating white blood cells from whole blood; breaking white blood cells to obtain broken white blood cells; and performing cell lysis and DNA-protein de-crosslinking on the broken white blood cells to obtain genomic DNA. By creatively breaking white blood cells and by breaking DNA and protein into small fragments, DNA-protein cross-linking sites are exposed to facilitate splitting and de-crosslinking, so that the extracted DNA is relatively more comprehensive and complete. The obtained DNA fragments satisfy the needs of subsequent database construction, and do not need to be broken again. Therefore, the method and kit specifically solves the problem that white blood cell fixation by preservation solution of a free DNA blood collection tube causes chromosome Z value abnormality, a large number of fragment deletions and repetitions.

Description

technical field [0001] The invention relates to the field of nucleic acids, in particular to a method and kit for extracting genomic DNA from whole blood. Background technique [0002] Among the newborns in my country every year, the proportion of birth defects is about 5.9%. The birth of children with defects has brought huge mental burden and economic pressure to the family; and with the increase of pregnant women's age, the prevalence of birth defects is significantly 70% of birth defects can be detected in advance. Compared with traditional prenatal screening methods such as serological screening and puncture diagnostic methods, non-invasive prenatal genetic testing based on the peripheral blood of pregnant women has the advantages of non-invasive, safe, accurate, and short detection cycle, and can effectively avoid invasive childbirth. Miscarriage and infection risks from pre-testing. [0003] NIPT detects the fetal genome by extracting plasma cell-free DNA, but the de...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 方楠白灵李红蕊王建伟伍启熹刘倩刘珂弟唐宇
Owner BEIJING USCI MEDICAL LAB CO LTD
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