Pig tissue chromatin immunoprecipitation processing method

An immunoprecipitation and processing method technology, applied in the field of animal immunology, can solve the problems of lack of systematic research methods, and achieve the effects of low cost, effective preparation and simple operation

Inactive Publication Date: 2017-10-17
HUAZHONG AGRI UNIV
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Problems solved by technology

There are many methods for studying 3D genomes, such as: 3-C (chromosome conformation capture), 4-C (chromosome conformation capture-on-chip), 5-C (chromosome conformation capture carbon copy), ChIP (chromatinimmunoprecipitation), ChIA-PET (chromatin interaction analysis by paired-endtag sequencing) and other technologies (BonevandCavalli 2016), and these technologies, especially ChIP, have been maturely applied in human and mouse cells and tissues, but there has been no systematic research on the ChIP technology in pigs. Research methods

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  • Pig tissue chromatin immunoprecipitation processing method
  • Pig tissue chromatin immunoprecipitation processing method
  • Pig tissue chromatin immunoprecipitation processing method

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Embodiment 1

[0038] A kind of porcine tissue chromatin immunoprecipitation (ChIP) processing method, comprises the pretreatment of tissue and doing chromatin immunoprecipitation in cells, i.e. ChIP, its feature comprises the following steps:

[0039] (1) Weigh 1g of fresh porcine tissue, place it in a 50mL centrifuge tube, and cut the porcine tissue into about 1-3mm 3 small pieces;

[0040] (2) Add 23.75 mL of 1% formaldehyde solution, treat at room temperature for 15 minutes, and carry out cross-linking under the condition that the tissue is fully shaken;

[0041] (3) Add 1.25mL of 2.5M glycine to a final concentration of 125mM to terminate crosslinking, treat at room temperature for 5min, continue to shake the tissue under crosslinking conditions, and after 5min, centrifuge at 2500rpm, 25°C for 5min;

[0042] (4) Discard the supernatant, take the precipitate, resuspend the precipitate with 20mL of compound phosphate buffer, blow it up and down with an electric pipette, wash it, and then...

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Abstract

The invention belongs to the technical field of livestock immunology, and concretely relates to a pig tissue chromatin immunoprecipitation processing method. The method is named as ChIP for short. The method specially aims at pig tissue for optimization design, employs a tissue lysate and a Dounce grinder for combination, and develops the method, the processes and the flows of pig tissue homogenate, the tissue can be fully cracked, the obtained chromatin amount is large, the quality is high, and the quality of a peak graph of the subsequent ChIP-Seq experiments is high. The method has the advantages of simpleness and rapidity, can effectively prepare a tissue suspension, can satisfy the core requirement of the ChIP-Seq experiments, and provides the effective approach for developing the pig tissue epigenomics researches.

Description

technical field [0001] The invention belongs to the technical field of livestock immunology, and in particular relates to a pig tissue chromatin immunoprecipitation (ChIP) processing method. Background technique [0002] At present, the research on 3D genome is becoming more and more popular, and histone modification has also become a hot spot, which plays a vital role in the regulation of gene expression. There are many methods for studying 3D genomes, such as: 3-C (chromosome conformation capture), 4-C (chromosome conformation capture-on-chip), 5-C (chromosome conformation capture carbon copy), ChIP (chromatinimmunoprecipitation), ChIA-PET (chromatin interaction analysis by paired-endtag sequencing) and other technologies (BonevandCavalli 2016), and these technologies, especially ChIP, have been maturely applied in human and mouse cells and tissues, but there has been no systematic research on the ChIP technology in pigs. Research methods. [0003] The ChIP method refers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6806C12Q2527/125C12Q2527/127
Inventor 曹建华任瑞敏黄英孙艳王昇赵书红
Owner HUAZHONG AGRI UNIV
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