Ultrasound breaking method for animal tissue in chromatin co-immunoprecipitation

A technology of co-immunoprecipitation and animal tissue, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, to achieve the effects of saving consumption, saving consumables, and reducing time consumption

Inactive Publication Date: 2014-08-06
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology solves problems associated with current methods used by researchers when studying animals' biological material. It allows scientists to study them faster without having to expend much effort or resources on developing new treatments.

Problems solved by technology

This patents describes various ways to study how different parts of biology work together differently when interacting through specific molecules called chromosomal imbalances. These interference may lead to diseases like cancer. Current researchers often focus on analyzing this phenomenon by developing new technologies based upon chemical reactions involving enzyme activity.

Method used

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  • Ultrasound breaking method for animal tissue in chromatin co-immunoprecipitation
  • Ultrasound breaking method for animal tissue in chromatin co-immunoprecipitation
  • Ultrasound breaking method for animal tissue in chromatin co-immunoprecipitation

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1 Ultrasonic disruption method of animal tissue in chromatin immunoprecipitation (taking chicken embryo as an example)

[0035] 1. ChIP experimental sample preparation

[0036] After the collected eggs were sterilized, they were placed in an incubator with a temperature of 37.8°C and a humidity of 60%, and the eggs were turned every 2 hours. After incubation until the 16th day, the heart tissue of the chicken embryos was collected and quickly frozen in liquid nitrogen.

[0037] 2. Exploration of ultrasonic crushing experimental conditions and result verification in the ChIP experimental process

[0038] The reagents used in the experiments were from ChIP unless otherwise noted. TM G Tissue Kit (Millipore, USA)

[0039] Take about 50ug of frozen chicken embryo heart tissue, thaw slowly on ice and chop into pieces → add 1ml 1:1 mixed collagenase (Sigma, C0130, USA), DPBS (Gibco, C14190, USA) solution, and digest the animal in a shaking water bath at 37°C Centr...

Embodiment 2

[0043] Example 2 Ultrasonic disruption method of animal tissue in chromatin immunoprecipitation (taking pig tissue as an example)

[0044] 1. ChIP experimental sample preparation

[0045] Heart tissue from 6-month-old pigs was collected and quickly frozen in liquid nitrogen.

[0046] 2. Exploration of ultrasonic crushing experimental conditions and result verification in the ChIP experimental process

[0047] The reagents used in the experiments were from ChIP unless otherwise noted. TM G Tissue Kit (Millipore, USA)

[0048] Take about 50ug of frozen pig heart tissue, thaw slowly on ice and chop into pieces → add 1ml of 1:1 mixed collagenase (Sigma, C0130, USA), DPBS (Gibco, C14190, USA) solution, and digest the animal tissue in a shaking water bath at 37°C 3.5h→4℃, centrifuge at 800g for 15min, remove supernatant→add 500ul TSS / PI, pipette repeatedly with pipette tip→4℃, centrifuge at 800g for 5min, remove supernatant→add 1ml1×phosphate buffer saline (PBS) / 1% Formaldehyde...

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Abstract

The invention provides an ultrasonic breaking method for animal tissue in chromatin co-immunoprecipitation. The ultrasound breaking method comprises the steps as follows: firstly, adding a mixture of a collagenase solution and a DPBS solution at a proportion of 1:1 into animal tissue for an experiment; shaking for digesting the animal tissue at the temperature of 37 DEG C; centrifugalizing; removing supernatant fluid; performing conventional ultrasound breaking. The ultrasound breaking method can better acquire a DNA-protein compound segment with a proper size, and a foundation is laid for the success of the whole ChIP experiment.

Description

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Claims

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Application Information

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Owner CHINA AGRI UNIV
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