DNA index library building method based on high throughput sequencing

A construction method and labeling technology, which is applied in the field of nucleic acid sequencing and high-throughput sequencing, can solve the problems of inability to mix and sequence a large number of samples, low PCR amplification efficiency, and small number of labels, so as to reduce sequencing costs and increase sequencing throughput. Quantity, the effect of improving efficiency

Active Publication Date: 2012-04-11
BGI TECH SOLUTIONS
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Problems solved by technology

[0004] 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 PCR index primer, as shown in Table 1), and the PCR amplification efficiency is not high

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  • DNA index library building method based on high throughput sequencing
  • DNA index library building method based on high throughput sequencing
  • DNA index library building method based on high throughput sequencing

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Embodiment Construction

[0341] Embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will understand that the following examples are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention.

[0342] according to Figure 4 Optimized DNA tag library construction process to construct DNA tag library

[0343] One aspect of the present invention provides a set of DNA tags, the DNA tags include or consist of the following: at least 5, or at least 10, of the DNA tags shown in Table 2 or DNA tags that differ therefrom by 1 base, or at least 15, or at least 20, or at least 25, or at least 30, or at least 35, or at least 40, or 45, or at least 50, or at least 55, or all 59,

[0344] The DNA tags preferably include at least DNA Index1-DNA Index5, or DNA Index6-DNA Index10, or DNA Index11-DNA Index15, or DNA Index16-DNA Index20, or DNA Index21-DNA Index25 of the 59 DNA t...

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Abstract

The invention provides unique index sequences with a length of 7bp. The index sequences can be respectively imported into a DNA(deoxyribonucleic acid) index library through adapter link and PCR (polymerase chain reaction). The invention provides a method for building the DNA index library by using the index sequences based on a solexa sequencing platform of the current illumina company, and the method is applied to solexa DNA sequencing and has the effects on improving the preparation efficiency of the DNA index library, increasing the sequencing throughput of the DNA samples and lowering thesolexa sequencing cost of a single sample.

Description

technical field [0001] The invention relates to the technical field of nucleic acid sequencing, in particular to the technical field of high-throughput sequencing. 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 [0002] The Solexa DNA sequencing platform provided by Illumina can simultaneously add four kinds of fluorescently labeled nucleotides in one reaction, using Sequencing By Synthesis (SBS), which requires less sample volume, high throughput, high Accuracy, has 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 A base at the 3' end of the ta...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C40B40/06C40B50/06
CPCC12Q1/6869
Inventor 章文蔚张艳艳于竞田方陈海燕龚梅花周妍
Owner BGI TECH SOLUTIONS
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