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A kit, method and application for detecting the interaction between protein and dna

A protein and sample technology, applied in the field of molecular biology, can solve the problems of inapplicability to a small number of source samples, high cost, and affecting the recovery rate of ChIP-DNA, and achieve the effect of high universal applicability and low cost

Active Publication Date: 2021-02-09
SHANGHAI JIAOTONG 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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  • A kit, method and application for detecting the interaction between protein and dna
  • A kit, method and application for detecting the interaction between protein and dna
  • A kit, method and application for detecting the interaction between protein and dna

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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 protein and DNA, which at least includes exogenous histone antigen. Also disclosed is the application of the above kit in detecting the interaction of a small amount of protein and DNA, and the required sample amount can be as low as 1000 cells. The invention also discloses a method for detecting the interaction between protein and DNA and its application. The technical solution provided by the invention reduces sample loss and improves the efficiency of immunoprecipitation reaction by supplementing the antigen corresponding to the histone interacting with DNA to be studied in a small amount of initial samples. It has the following advantages: 1. The minimum required sample volume is 1000 cells, which is more suitable for the detection and analysis of the interaction between protein and DNA from a small number of samples; 2. ChIP experiments do not require specific kits and experimental equipment, and the cost is low. 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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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68C12N15/10
CPCC12N15/1013G01N33/68
Inventor 康亚妮赵小东徐伟毛湛睿
Owner SHANGHAI JIAOTONG UNIV
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