Method for preparing chromatin co-immunoprecipitation library by using trace of clinical puncture sample and application of method

Abstract

The invention discloses a method for preparing a chromatin co-immunoprecipitation library by using a trace clinical puncture sample and an application of the method. The invention successfully develops the method for preparing the chromatin co-immunoprecipitation sequencing library based on trace of clinical puncture sample by utilizing high-sensitivity Tn5 transposase, and ChIP-seq library construction can be effectively and rapidly carried out on the trace of puncture tissue sample. Compared with a traditional ChIP-seq library building mode, the method has the advantages that the initial sample amount is greatly reduced (5-50 mg), the library building time is effectively shortened (about 1.5 days), and the requirements of the sequencing market on trace samples and large-scale sample processing speed are met.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing a chromatin immunoco-precipitation library using a small amount of clinical puncture samples and an application thereof. Background technique [0002] Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is a revolutionary technology that combines chromatin immunoprecipitation (ChIP) with high-throughput sequencing. This technique can identify binding sites on chromatin, such as transcription factors, histone modifications, nucleosomes, and RNA-binding proteins, on a genome-wide scale. Different from the traditional chip-based ChIP-chip (chromatin immunoprecipitation combined with DNA tiling arrays), ChIP-seq provides a new strategy for studying protein-DNA interactions with high resolution, high specificity, and high coverage. ChIP-seq technology can be applied to any species whose genome sequence is known, can study the interaction bet...

AI Technical Summary

Problems solved by technology

[0003] Although the traditional ChIP-seq technology has the advantages of high resolution, low noise, and high coverage, there are still some problems: First, the current ChIP-seq requires too much sample input, and the total amount of tissue samples needs to be at least 200mg Second, the chromatin immunoprecipitation experiment steps are cumbersome in the early stage, requiring formaldehyde cross-linking, cell lysis, ultrasonic disruption, antibody enrichment of target protein and DNA complexes, target protein digestion and cross-linking, purification of target DNA fragments, and DNA fragment end repair, target DNA fragment end adenylation, target DNA fragment connection adapter, PCR amplification and other steps

The cumbersomeness of the steps leads to defects such as long library construction cycle and low success rate

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