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Identification method of BPDE adduct gene

An identification method and gene technology, applied in the field of identification of BPDE adducted genes, can solve the problems of high-throughput sequencing false positives, information loss, etc.

Active Publication Date: 2019-07-30
SHANXI MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0006] In this technical method, due to differences in the state of cell culture and the state of cells after exposure, and the ultrasonic fragmentation of each sample cannot ensure that each The exact same fragment size is obtained every time, and some genes at the binding sites may be interrupted during ultrasound, resulting in information loss. In addition, there are some false positive results in high-throughput sequencing

Method used

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  • Identification method of BPDE adduct gene
  • Identification method of BPDE adduct gene
  • Identification method of BPDE adduct gene

Examples

Experimental program
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Embodiment

[0083] A kind of identification method of BPDE addition gene, comprises the following steps:

[0084] 1) Observe under the microscope that the degree of cell confluence in the culture medium grows to 70%-80%, and the cells are in good condition at this time.

[0085] In this step, confluency is a measure of the density of cells in the dish. This indicator is a well-known concept in the art. Only when the cell confluence reaches 70%-80%, it means that the cell is in good condition, then proceed to the following steps.

[0086] 2) Add formaldehyde to the medium to fix the cells in the formaldehyde, vortex the culture dish to mix the formaldehyde in the medium, and let stand at room temperature for 10 minutes.

[0087] In this example, the preferred method of adding formaldehyde is to add methanol solution (the solvent is deionized water), and the preferred concentration of formaldehyde solution is: add 275 μL of 37wt% formaldehyde solution per 10 mL of medium. Try to avoid di...

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Abstract

The invention relates to the field of identification of BPDE adduct genes, in particular to an identification method of a BPDE adduct gene. Based on the principle of chromatin immunoprecipitation (ChIP), the BPDE-DNA adduct enriched by Anti-BPDE antibody is sequenced with high throughput to construct B [a] P adduct target gene site accurate localization technique. According to the method, the detected BPDE adduct target gene is a direct action between the BPDE and the gene, and small molecules do not need to be chemically modified in the detection process, so that the original activity of thesmall molecules is not influenced, the method is not limited by any cell and tissue type, pure DNA is not required to be used, even a whole cell lysate can be used, and the method can be widely applied to the identification of the small molecule adduct target gene.

Description

technical field [0001] The invention relates to the identification field of BPDE adducting genes, in particular to a method for identifying BPDE adducting genes. This method can directly identify target genes that can be added to BPDE in chromatin. The previously reported role of B[a]P or metabolite BPDE at the genome level is mainly based on the expression difference of one or some genes. The relevant upstream or downstream genes may play a certain biological function. In this method, the detected BPDE adducted target gene is the direct interaction between BPDE and the gene, and there is no need for small Chemical modification of molecules will not affect the original activity of small molecules, not limited by any cell and tissue type, does not require the use of pure DNA, and even whole cell lysates can be used, which can be widely used in the addition of small molecules to target genes Identification. Background technique [0002] Polycyclic aromatic hydrocarbons (PAHs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2535/122C12Q2545/113
Inventor 郑金平曹彬吕懿王丹穆箭兵
Owner SHANXI MEDICAL UNIV
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