Method for rapidly preparing chromatin immunoprecipitation sample

Abstract

The invention discloses a method for rapidly preparing a chromatin immunoprecipitation sample. The method comprises the following steps: performing formaldehyde crosslinking on a cell to fix a complex of a protein and a nucleic acid, extracting the complex of the protein and the nucleic acid, and breaking a chromatin segment into small segments in an ultrasonic or enzymatic hydrolysis manner; suspending an antibody onto a solid-phase support, i.e. a magnetic bead, adding the obtained small segments onto the prepared magnetic bead, performing incubation to obtain a magnetic bead antigen antibody complex, washing an uncombined substance away after incubation is finished, and resuspending the magnetic bead antigen antibody complex to obtain a magnetic bead antigen antibody complex sample to be detected. The method is easy to operate; the prepared magnetic bead antigen antibody complex sample can be directly used for PCR detection or amplification and construction of a DNA library, so that the sample preparation time is obviously shortened, meanwhile, the shortcoming of weaker enrichment signals of certain proteins and DNAs caused by excessive elution steps is overcome, the detection sensitivity is greatly improved, and influence on subsequent PCR identification and DNA segment amplification is avoided.

Description

technical field [0001] The invention belongs to the technical field of chromatin immunoprecipitation (ChIP), and more specifically relates to a rapid preparation method for chromatin immunoprecipitation (ChIP) samples. Background technique [0002] Histone is a basic protein in the chromatin of eukaryotic cells. It contains a lot of basic amino acids such as arginine and lysine, which add up to about 1 / 4 of all amino acid residues. Histones bind to the negatively charged double-helix DNA to form a DNA-histone complex. Histones are the basic structural proteins of eukaryotic chromosomes. They are a type of small molecular basic proteins. There are five types: H1, H2A, H2B, H3 and H4 histones. They are rich in positively charged basic amino acids and can Interacts with negatively charged phosphate groups in DNA. [0003] Various post-translational modifications (PTMs) (i.e., methylation, acetylation, ubiquitination, and sumoylation) of many histones occur at lysine sites. A...

AI Technical Summary

Problems solved by technology

[0008] The preparation results of chromatin immunoprecipitation samples play an important role in subsequent sequencing analysis, and its preparation efficiency also directly affects the time of the entire ChIP-Seq operation. However, the existing sample preparation process is often time-consuming and costly. The detection sensitivity is also low

Images