Preparation method of accurate quantitative ATAC-seq library and kit

Abstract

The invention provides a preparation method of an accurate quantitative ATAC-seq library and a kit. The kit comprises two kinds of linkers, wherein the first linker is 5'-S1-S2-S3-S4-3', and the second linker is 5'-S5-S6-S7-S8-3'; S1 is a first sequencing primer; S2 is a random tag sequence; basic groups of S2 are one or a random combination of more of the sequences; the number of the basic groupsof S2 is any integer of 1-95; S3 is a fixed sequence; the basic group sequences of S4 and S8 are the same and are Tn5 transposon fixed sequences; S5 is a second sequencing primer; S6 is a random tagsequence; basic groups of S6 are one or a random combination of more of the sequences; the number of the basic groups of S6 is any integer of 1-100; S7 is another fixed sequence; the two kinds of linkers and Tn5 enzyme are incubated to obtain a transposon complex; then the transposon complex and a test sample are incubated to obtain an amplification template; and the amplification template is subjected to PCR amplification to obtain a sequencing library.

Description

technical field [0001] The present invention relates to the field of biotechnology, more specifically, to an accurate and quantitative ATAC-seq library preparation method and kit. Background technique [0002] At present, we can use the transposase accessible chromosome (ATAC) assay to study open chromatin regions in the genome, which are important binding sites for transcription factors and transcription elements. The methods currently used to study open chromatin regions are: 1. DNase-seq, the naked DNA fragments are cut with DNaseI endonuclease, the cut DNA fragments are sequenced, and compared with the known whole genome sequence, You can find out which part of the cut-off region is, and you can find the developed chromatin region, but this method is cumbersome and takes a long time (2-3 days), and requires a large number of nuclei for experiments (more than one million nuclei), And the repeatability is also poor. 2. ATAC-seq (Assay for Transposase-Accessible Chromatin...

AI Technical Summary

Problems solved by technology

These kits can satisfy different sources of plant samples, and satisfy scientific research applications to the greatest extent. However, there is currently no method for accurate and quantitative library preparation on the market. The experimental process of the library construction method in the conventional ATAC-seq library There are still systematic errors, especially for a small number of samples, it is necessary to increase the amount of sequencing to improve the accuracy of detection, and in this case the repetition rate of data will increase significantly, which limits the application of this method in precision medicine and disease prediction other needs

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