Method for introducing molecular tag in library construction suitable for Illumina next-generation sequencing platform, linker sequence and application thereof

Abstract

The invention provides a method for introducing a molecular tag in library construction suitable for an Illumina next-generation sequencing platform, a linker sequence and a design method, a method for constructing a library by using the linker sequence, and a kit containing the linker sequence. Wherein the linker sequence comprises 5'-P-E-F-Q-3', P is a sequencing platform pairing sequence, E isa random base sequence composed of the same base, F is a given base sequence, and Q is a Tn5 transposase terminal recognition sequence. The linker sequence and the library construction method providedby the invention can successfully construct the library for DNA samples as low as 50 pg, and are suitable for preparing DNA library construction kits and disease diagnosis kits.

Description

technical field [0001] The present invention relates to a high-throughput sequencing library construction method, in particular to a library suitable for the Illumina next-generation sequencing platform, especially a method for introducing molecular tags in library construction, a linker sequence, a linker sequence design method, and applications. Background technique [0002] Next-generation sequencing is also called NGS sequencing or high-throughput sequencing. As the name suggests, high-throughput sequencing has a high throughput of sequencing reactions. We can get 1Gb to 3Tb of sequencing data in one sequencing reaction. The human genome includes 20,000 genes encoded by it. The remaining gene sequences can be obtained at one time. The accuracy of high-throughput sequencing is also very high, with a single-base accuracy exceeding 99.9%. The high-throughput sequencing platform has achieved breakthroughs from scientific research to clinical application, and is widely used ...

AI Technical Summary

Problems solved by technology

The traditional library construction method is cumbersome and time-consuming. During the PCR amplification process, a large number of copy duplications will be randomly generated. When calculating the mutation frequency of gene mutation sites, large errors are likely to be caused by repeated sequence comparisons, and the The initial DNA quality requirements for library construction are relatively high (generally above 50ng)

Images