Tag library constructing method based on DNA (deoxyribonucleic acid) adapter connection as well as used tag and tag adapter

Abstract

According to the invention, based on a DNA (deoxyribonucleic acid) tag library preparation method provided by a solexa sequencing platform of the illumine corporation, a special tag sequence with length of 6 bp is designed, the tags are embedded into DNA adapters, and the DNA adapters are connected to introduce the tag sequence, thus a library construction method of a DNA tag library is successfully constructed, and the constructed DNA tag library can be applied to solexa DNA sequencing.

Description

technical field The invention relates to the technical field of nucleic acid sequencing, in particular to a method for preparing a DNA tag library. In addition, the present invention also relates to labeling technology and a method for constructing a label library in the same reaction system for multiple samples. The method of the present invention is particularly suitable for the second generation sequencing technology, especially the solexa sequencing technology. Background technique The Solexa DNA sequencing platform provided by Illumina can simultaneously add four kinds of fluorescently labeled nucleotides in one reaction, and adopts Sequencing By Synthesis (SBS), which requires less sample volume and high throughput. High accuracy, with the characteristics of simple and easy-to-operate automation platform and powerful functions [1-4]. Library construction first needs to carry out end repair on the target fragment, connect the "A" base at the 3' end of the target fragm...

AI Technical Summary

Problems solved by technology

However, there are some defects in the method for preparing the tag library provided by Illumina: First, at present, Illumina only provides 12 tag sequences with a length of 6 bp, and the number of tags is small. Mixed sequencing of samples will be a huge defect; second, the current label library construction method provided by Illumina is to import the label sequence into the target fragment library through PCR reaction, which requires 3 PCR primers to amplify the target fragment (two Common primers and a label PCR primer, as shown in Table 1), and the PCR amplification efficiency is not high

Third, the linker in the label library construction method provided by illumina does not contain the label sequence. Each label library needs to pass a PCR reaction to import the label sequence, and then for each label library, it needs to be cut and recovered, and then the glue is recovered. It is not only time-consuming and labor-intensive, but also expensive

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