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Kit and method for detecting interaction between proteins and DNA, and applications of kit and mehod

A kit and protein technology, applied in the field of molecular biology, can solve the problems of high cost, unsuitable for a small amount of source samples, affecting the recovery rate of ChIP-DNA, etc., and achieve the effect of low cost and high universal applicability

Active Publication Date: 2018-10-19
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, the addition of RNA will affect the recovery rate of ChIP-DNA obtained during subsequent removal, thus reducing the enrichment effect of ChIP to a certain extent.
[0005] There are two main disadvantages of the prior art: 1. The minimum sample input quantity required is still a relatively large number of cells, which is still not suitable for ChIP analysis of a small amount of source samples; 2. The experimental process involves commercial kits and complete Automated sample handling systems, which are expensive and not universally applicable or versatile

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  • Kit and method for detecting interaction between proteins and DNA, and applications of kit and mehod
  • Kit and method for detecting interaction between proteins and DNA, and applications of kit and mehod
  • Kit and method for detecting interaction between proteins and DNA, and applications of kit and mehod

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Embodiment Construction

[0030] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental materials and reagents used in the following examples can be obtained from commercially available channels unless otherwise specified.

[0031] In this embodiment, the method for detecting the interaction between protein and DNA specifically includes the following steps:

[0032] 1. Wash the K562 cells cultured in a 10cm dish with 2mL PBS twice;

[0033] 2. Formaldehyde cross-linking: add 37% formaldehyde to 5ml medium to make the formaldehyde concentration of the medium finally 1%, shake at room temperature at 100rpm for 10min;

[0034] 3. Termination of fixation: Add 2M glycine solution to the cross-linking buffer to make the final concentration 0.125M, mix well, shake at 100rpm, room temperature for 5min...

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Abstract

The invention discloses a kit for detecting the interaction between proteins and DNA, wherein the kit at least includes an exogenous histone antigen. The invention also discloses an application of thekit in detecting the interaction between a small number of proteins and the DNA, and the required sample amount can be as low as 1000 cells. The invention also discloses a method for detecting the interaction between the proteins and the DNA and an application thereof. According to the technical scheme provided by the invention, sample loss is reduced and the efficiency of immunoprecipitation reaction is improved by additionally adding a corresponding antigen of histone requiring to be researched and interacting with DNA into a small number of initial samples. The kit has the following advantages: 1, the minimum required sample number is 1000 cells, and the kit is more suitable for detection and analysis of the interaction of a small number of proteins and DNA from the samples; and 2, a ChIP experiment does not require specific kits and experimental equipment, the cost is low, and the kit has high universal applicability.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a kit, method and application for detecting the interaction between protein and DNA. Background technique [0002] The interaction between protein and DNA plays a vital role in the regulation of gene expression and the formation of chromosomes. The combination of transcription factors and specific DNA sequences is an important mechanism for the regulation of specific gene expression. Expressions such as histone methylation Epigenetic modification also plays a very important role in gene expression, so studying the interaction between proteins and DNA can better help people understand complex biological processes. Chromatin immunoprecipitation (Chromatin Immunoprecipitation, ChIP) is a commonly used technique for studying the interaction between proteins and DNA. It is based on the principle of enriching DNA bound to specific proteins. On this basis, ChIP-qPCR, ChIP -ch...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12N15/10
CPCC12N15/1013G01N33/68
Inventor 康亚妮赵小东徐伟毛湛睿
Owner SHANGHAI JIAO TONG UNIV
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