Epigenetic DAP-seq sequencing library building method

Abstract

The invention belongs to the field of high-throughput sequencing library building, and particularly relates to an epigenetic DAP-seq sequencing library building method. The sequencing library buildingmethod provided by the invention mainly comprises the processes of vector building, TF in-vitro expression pre-experiment, DAP-seq library building and the like. According to the sequencing library building method provided by the invention, the genome DNA extracted from cells can be directly combined with target TF, so that genome DNA modification can be reserved, and compared with the prior art,the sequencing library building method is closer to an in-vivo test.

Description

technical field [0001] The invention belongs to high-throughput sequencing library construction technology, in particular to an epigenetic DAP-seq sequencing library construction method. Background technique [0002] In epigenetics research, the discovery of transcription factor (TF, Transcription factors) binding sites has always been a difficult problem, and CHIP-seq is the most effective technique to directly study the target TF binding site. CHIP-seq uses specific TF antibodies to pull out the DNA fragments that bind to TF through co-immunoprecipitation, and then through high-throughput sequencing, it can directly detect the binding sites of specific TFs on the genome and analyze the binding sites. motif (signature sequence of binding site). [0003] However, CHIP-seq is not applicable to all species, and there are mainly the following difficulties: (1) Difficulties in antibody preparation: For popular transcription factors in common model species such as humans and mic...

AI Technical Summary

Problems solved by technology

This scheme still has a shortcoming: for some species, the transgenic experiment is very difficult or the cycle is very long, and the experimental threshold is still very high

The biggest shortcoming of this method is that the DNA is artificially synthesized, and there is no modification information (such as DNA methylation) on the original genomic DNA

However, information such as DNA methylation is an important factor affecting TF binding, so methods such as SELEX and PBM are difficult to restore the real binding situation of TF in vivo, and there is still a big difference in the effect of CHIP-seq.

[0005] It can be seen that the existing technology generally has the disadvantages of poor effect, destroying the modification information on the original genomic DNA, and making it difficult to restore the real situation of TF in vivo

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